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Hi friends,
So I'm new to cell culturing research and synthetic biology and I'm working with MDA-MB-231 cells. Fairly regularly I have to at least split plates in order to maintain the cells, but I am also passaging cells for other things. Normally on a 10cm petri dish I'd plan to seed ~5.0E5 cells/mL for regular splitting and in a 96 well plate I'd seed ~4.5E4 cells/mL for my experiments. One thing I'm confused on still is using the hemocytometer to calculate cell density because I'm not entirely sure what my dilution factor should be.
Normally, I'll remove the old media, wash with PBS then trypsinize cells, after incubating for 5mins I add a bit more media until the total volume is 10mL, then I spin everything down. After that, I aspirate out the supernatant and am left with a small pellet of cells that I then break up and dilute with ~2-6mL of fresh media before using the hemocytometer to count the number of cells in 10uL.
My question is since I'm removing all the old media after centrifugation but then am adding new (different amount) media prior to counting, how does that impact the overall dilution factor?
Here's an example of what I was taught to do to find the amount of cells in 1mL, I just wanted to confirm that I'm on the right track. I was told if I resuspended my pellet in 3mL of media and then counted 75 cells I'd have:
75 cells/10E-4mL media
75 E4 cells/1mL media
7.5E5 cells/1mL media --> multiply this by 3 since you used 3mL to dilute before counting to get total number of cells (is this what accounts for the dilution factor?)
Total of 2.25E-6 cells that could be split however you see fit? Sometimes determining how many plates to get out can feel like a guessing game. Let me know if you see any issues with my method or suggestions that might be helpful, I welcome them. Please do try to be kind as I'm still learning and figuring things out. Thank you.
Kylie
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“One thing I'm confused on still is using the hemocytometer to calculate cell density because I'm not entirely sure what my dilution factor should be.”
The dilution factor that you need to take into consideration is when you dilute the 10ul cell suspension with trypan blue for the cell count using hemocytometer.
When you take 10ul of cell suspension to count using hemocytometer, you should add 10ul of trypan blue. This will give you 1:1 dilution. So, the dilution factor will be 2.
Suppose your cell count using hemocytometer is 75 (as you mentioned).
Then your cell count per ml = 75 x 10^4 x dilution factor (2).
So, cell count/ml= 150 x 10^4 cells/ml
Since you have resuspended the pellet in 3ml of media, you will have to multiply the above cell count by 3 to get the total number of cells.
So total number of cells in 3ml media = 150 x 10^4 x 3 = 450 x 10^4 = 4.5 X 10^6 cells.
Total of 2.25E-6 cells is not correct because you have not multiplied this count (2.25E-6) by the dilution factor which is 2. (The dilution factor of 2 comes from the 1:1 dilution of cell suspension with trypan blue for cell counting using hemocytometer).
So, if you have 4.5 x 10^6 cells in stock, you could calculate how many 10 cm petri dishes or 96 well plate you could seed. For instance, you could seed nine 10cm petri dishes from 4.5 x 10^6 cells, if you are planning to seed ~5.0E5 cells per 10 cm petri dish.
Best Wishes.
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Researchers have discovered that breast cancer cells are more likely to jump into the blood when people are resting. The revelation is relevant because cancer is at its deadliest when a tumour’s cells worm their way into the bloodstream and travel to a new location in the body to set up shop — a process called metastasis. It doesn’t mean that people with breast cancer should avoid sleep — that’s bad for you. But it does give clinicians a better understanding of how to track these rogue cells. “The first lesson for me is that the time of day you take a blood sample can give you misleading information”, says cancer biologist Chi Van Dang...
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I am analysing several drugs on tumour cell lines. I need a quick way to calculate IC50 based on data generated.
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Dear researcher
Sigma plot software is recommended.
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In case of mTOR I notice cancer research wants to block this pathway right? So would resistance training, amino acids and insuline be dangerous to some extent? Someone with genes for cancer growth can he or she also have more danger with training?
Or is it only when high doses of insulin, growth hormon and such are ingested or injected?
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Interesting
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Could you please provide any tips or ideas you have for creating the finest meta analysis review on cancer? Rather than writing a review article, I'd like to begin working on a meta analysis.
regards
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I totally agree with Timo Van Canegem that the word by itself "cancer" is such a broad term and I would narrow down to a particular sub-type and/or treatment. My best suggestion for this would be if you are uncertain as to which type to focus on to look at what type or types are treated or investigated in your organisation. For example, where I work currently, they are particularly interested in lung cancers and where I was working before has a special focus on bowel and breast cancers.
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We want to know if one protein can be used as a breast cancer biomarker or not. So we'll have to compare normal and patients, and our question is whether we can get samples from patients who received chemotherapy before?
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thank you for answring my quesion dear Dr Erum Zafar
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I should use the BCH for inhibition experiment for LAT1. I saw that BCH is soluble in 1M NH4OH but I cannot use this solution with cells.
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The following attachment may be relevant to this query
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in cancer therapy, sometimes you have to target multi-targets to understand the molecular pathway of the specific protein. CHyMErA can target multi targets but it is not safe and has a high risk of extra mutation!
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thank you dear Dr Murali Mohana Rao Singuru
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Is there a chance for cancer research fields, such as exponential growth of infected blood cells methodology, to be related to COVID-19 research generally?
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Have a look at this useful RG link.
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This seminal paper by Paul Mischel rediscovered the relationship between extrachromosal DNA (ecDNA) and cancer:
1. How much would it cost to replicate the part of the experiment that generated subcutaneous tumors from seeds containing 200/2000/20000 cells of FACS-sorted EGFRvIII High/EGFRvIII Low subpopulations?
2. How much would it cost to sequence these tumor cells?
Thanks for your help!
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Shalini Sanyal Thanks for your answe.
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Hi
I'm trying to design a simple cube-shaped DNA origami with specific dimensions from just single stranded DNA oligos. 
Since there is no specific scaffold the origami is based on, cadnano is difficult to use...
I understand you can use Nupack... but I'm not sure how to create 3d objects using Nupack. 
Would there be a better program fit for this or what would be the ways doing so in nupack?
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Hi, I hope you already found an answer to this, but I think for cubes and other structures with specific dimensions ADENITA is a good option. Adenita SAMSON edition is a SAMSON (https://www.samson-connect.net/) plugin. you only need to add the app in the shop. You can use freeware option of SAMSON and it works quite nice.
De Llano, E.; Miao, H.; Ahmadi, Y.; Wilson, A.J.; Beeby, M.; Viola, I.; Barisic, I. Adenita: interactive 3D modelling and visualization of DNA nanostructures. Nucleic Acids Res. 2020, 48, 8269–8275, doi:10.1093/nar/gkaa593.
Best
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I want DNA microarray cancer data sets with features and samples to form a gene expression matrix for applying intelligence techniques (such as basiayan neural network algorithm) to classify cancer data sets. Please help me.
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Try to have a look at two publicly accessible warehouses: arrayExpress and GEO and you can search for experiments designed for cancer studies.
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Permissible limit of arsenic in drinking water was 50 μg/l by WHO which was later reduced to 10 μg/l. Currently many countries like Bangladesh and China have not updated their own Permissible limits. However, this is been observed that on the event of long term exposure of arsenic at low concentration that is less then 10 μg/l in drinking water, health complication arises.
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I made a stock solution (25mM) of Pioglitazone hydrochloride (Tocris) in DMSO. It has dissolved finely. Then I treated the cells maintained in serum free DMEM with a final concentration of 100 micromoler of Pioglitazone. Here it forms crystals, precipitates in the culture vessel, but I am not getting the desired effect of it on the cells. Please suggest how to achieve the proper effect of Pioglitazone treatment?
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I don't have a solution, bur we also see hair-like crystals forming in our culture media when pioglitazone concentration ≥10µM.
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Cell culture.
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It is about 6-12 hours, it grows really fast. For me, A flask is fully confluent after 48 hours, even it is splitted at 1-5.
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There is an increasing amount of research on the ketogenic diet in cancer therapy. However, I can't find much about this diet in those guidelines produced by governmental cancer research institutes such as American Institute for Cancer Research (AICR). As a matter of fact it seems like the word ketogenic is totally missing from the databases.
Am I missing something?
Sincerely Yours,
Christer Sundqvist
PhD Helsinki
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Thanks for the answers here
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Gene expression in the liver is zonated , is during aging a shift in zonation possible of the expressed genes?
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I think it is...
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The main ‘party trick’ of cancer stem cell (CSC) seem to be that it provide another explanation for tumour heterogeneity (i.e. cell divide asymmetrically give rise to progeny cells that are different). Secondly, asymmetric cell division is what makes this model ‘useful’ (i.e. clinically relevant): CSC model refer to a subpopulation of cells that must be targeted for effective thematic outcome. But if asymmetric cell division does not take place (i.e. if all cells divide symmetrically) than any therapy that kill cancer is in fact targeting the CSC (i.e. there is no point in identifying CSC- each cell would be one under symmetric cell division).
So, is asymmetric cell division a hallmark of CSC model? Any good literature on this?
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Hi!
I am trying to find a good immunocompetent mouse model to study triple negative breast cancer (TNBC). The 4T1-BALB/c model is perhaps not ideal for studies on immunosuppression and as 4T1 is derived from BALB/c mice, it cannot be transplanted in the C57BL/6 strain. Can someone suggest a good animal model to study TNBC-induced immunosuppression?
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Suhnrita Chaudhuri E0771 in C57BL/6 mice is, but still controversial as a pure TNBC model
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Hi,
I have been injecting AML primary cells in NGS-SGM3 mice and they developed leukemia. After recovering the human AML leukemic cells from the bone marrow it is difficult for me to keep them in culture and especially to expand them. I am using SFEM media with CC100 cytokine cocktails + Tpo. This media worked well for primary AML but is not giving me good results with the same cells isolated from the mice/xenograft models.
How can I optimize the condition to maintain AML cells recovered from the mice?
Thanks
Patrizia
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Did you ever find an answer for this? I am especially curious for the application of Cas9 editing in which the cells should be dividing to have highest efficiency (not just surviving but expanding)
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I'm using the BrdU antibody of the roche kit but i can't see any signal.
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In this case, it might be better to use another proliferation marker such as EDU because BrdU staining is mainly depending on denaturation by Hcl or you could try heating instead of Hcl treatment.
Good
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I had a contaminated cell culture I suspected several things, including the media. I put the media (only) in a T-25 flask and used 2 agar plates to check bacterial contamination (one opened in the laminar air flow the other in the CO2 incubator) and left them is in the CO2 incubator at 37 oC (after I cleaned it),
two days later I didn't find any thing on the agar plates and no changes in color or clarity of the media in flask however I examined the flask under the microscope and found these things .. What could it be?
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I understand that this can be very frustrating. The other answers are excellent responses for your problem. It is interesting that you had no contamination in your T-25 or agar plates.
I have found that without a cell layer to guide you, it is quite difficult to locate the media in a flask. Are you absolutely sure that you were looking at the media and not the bottom of the flask - the patterns in your picture resemble scratches and divots on the outside of the flask.
In any case, to help prevent contamination in your media, you can aliquot into 50 ml tubes and use those instead of an entire bottle at the same time.
Hope this helps!
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I am working with rectal swab samples in the field of colorectal cancer research. Does anyone have experience (or can point towards any relevant literature) of specifically working with proteins (extraction, characterization, or anything even remotely connected to protein work) in rectal swab samples (or any mucosal swab samples).
Thank you!
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Hi Colleague
Find the paper at the following URL may help you:
Regards...
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I just asking cancer expertise to suggestions the best books can read by MSc students before start their PhD program in cancer research
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Hallmarks of Cancer: The next generation
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I'm fairly new to cancer research and there's a host of articles on Sunitinib being administered via oral gavage in mice models but I'm unable to find any information on studies that have explored directly injecting tumors with Sunitinib. Just curious.
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Hi Paul
The name of the article is as follows:
Injecting activator of a powerful tumor suppressor directly into the cancer increases tumor destruction, decreases toxicity. Science Daily May 23, 2017.
Regards.
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Hello Everyone,
I've been writing a review article (almost done) on cancer research. Is anyone interested in collaborative writing and publication? Please let me know. We can share our ideas on extending the content of the paper and get it published mutually.
Professionals with an interest in cancer research are most welcome.
Thanks,
Dr. Ranbir
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I am now writing the article on the relationship between susceptibility genes and dysbiosis.
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I currently doing a research for anticancer drugs and have a plan to use CA 15-3 as tumor marker to measure the effectivity. Some reference tell that CA 15-3 and MUC 1 are synonym, but i also found some reference that stated CA 15-3 and MUC are different. My question is "Could I use ELISA kit for MUC 1 to measure the CA 15-3?" because my local supplier can't find the ELISA kit for CA 15-3 in affordable price.
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In principle yes, but with slight modification on the recommended protocol
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I read many papers about exosome isolation from fresh blood or frozen plasma, but not for frozen whole blood. I am wondering whether can I get exosomes from cryopreserved whole blood? One of my workmate told me the blood cells were broken if crypreserved and would affect the isolation of exosomes? What I need is the exosomes and their surface biomarkers. Could anyone give me your prescious advices? thank you so much.
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yes that can be done because DMSO has helped maintain the cells intact without any physiological disruption.
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Could anybody please tell me what are the different types of cells that originate from HNSCC patient samples. What are the type of studies that it can be used for?
I understand there are epithelial, fibroblasts, and keratinocytes that can be derived from HNSCC tumor samples. Can immunological studies be done using these type of cells?
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It really depends on what type of immunological studies you intend to run. Going back to the lymphocyte example I might do it in a deferent way .. by bringing lymphocyte form a different source and incubate the lymphocyte with the HNSCC and look for a response.
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I am looking for a scientifically oriented internet site providing new research in the category of cancer treatments.
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Thanks
Timothé Ménard
!
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I isolated a large amount (I hope) of exosomes and I need to quantify the membrane proteins with a Bradford assay before doing a WB.  I started off with 70 ml of conditioned medium which I managed to concentrate to about 1 ml before ultracentrifugation. After ultracentrifugation I added 200 ul PBS to my samples. My expectation is that the concentration of exosomes is quite high. So now I need to do a Bradford assay and  I would like to know how to dilute my samples.
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I'd like to kindly confirm from your response if the appropriate assay type for exosomal protein quantification with nanodrop is 1A/cm = 1 mg/ml.
I understand this is a general reference setting for protein solution, but I'd like to ask if IgG assay type is also relevant considering the lower detection limit which is about 0.06mg/ml compared to 0.08mg/ml for 1A/cm = 1 mg/ml assay type.
Thanks!
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Hello,
I am new to cancer research and I am trying to optimize my small cell lung cancer cell protocol.
I purchased a cell line from ATCC (H1105) and I have been growing the cells in RPMI 1640 media (10% FBS + penicillin/streptomycin.
I am looking for anyone who was worked with these cells in the past and who can let me know if these cells look normal?
I noticed the cells grow very slowly and form spheres.
Am I using the right media? How can I get the cells to grow faster? Can someone refer me to a good SCLC cell culture protocol? Thank you.
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Simple mechanical mixing is fine especially since your cells haven't formed confluent clusters yet.
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I need to confirm a knockdown of HER2 in SKBR3 and BT474 cells, have had problems blotting for HER2 in the past so any advice with regards to the protocol or antobody choice is much appreciated, thanks
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Thank you for all your amazing work! All the best!
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For my cancer research, I am doing cell culture with HPNE and MIA PACA. So, I would like to know the answer.
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Thank you
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I have transfected GFP-AR in to HEK293 and LncAP cell lines in serum free medium (HEK293-DMEM and LncAp-RPMI1640) using Lipofectamine. After how long should i change the medium? and should change it with serum free medium or serum containing medium (10% FBS)? 
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1. Transfect cells with media contain plasmid and transfection reagent without serum and antibiotics.
2. Allow at least 6 hours transfection (10-16 hrs is better).
3. Change the media and add regular complete media (contain FBS and antibiotics)
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I am studying on cancer research and ı used bcl 2 antibody to understand there is apoptosis or not. I got good band ı tested with ponceu red. Finally I got this result.ı used uvp chemi doc it2 imager to preview membrane.I tried a lot of times what do you think about this result?
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Hi Berna,
As Dr Mohammed comments, we need more details to understand the problem and think about how to help you. For example, may you show us the SDS-PAGE before blotting in order to analysis if the amount of protein loaded is adequated or to much, as it appears? The acrylamida percentage? Molecular weight of your target proteins? Previous treatments of the proteins? And finally, it is neccessary add a lane with molecular weight markers.
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MDA-MB-231, thawing
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From my personal experience, it seems like there are three issues that might have been contributing to your problem:
1- Not diluting the DMSO after thawing. You can fix this by adding about 5X the volume of cells of cell culture media (if you have about 2 mls of cells, add it to 10 mls of media). Then, you should centrifuge to pallet your cells and aspirate as much of the DMSO+media mix as possible after centrifugation. Finally, you can add more media and transfer the cells to a cell culture dish. This should get rid of almost all of the DMSO and give the cells a "happy" environment.
2 - Trying to change the media too soon. Cells can take quite some time to attach, especially after the stress of freeze/thaw. Six hours does not seem like enough time to change the media. Moreover, they might secrete factors in the media that will aid their growth and proliferation, and by changing the media too soon you get rid of those factors. Wait 24 h before changing the media: you will probably see a lot of (dead) cells floating, which is normal. Just aspirate the media and add more. After this, you should see some cells happily attached. Do not change the media or split cells before they are about 60% confluent (which can take 5-7 days for MDA-MB-231, depending on how many survive thawing), unless there are a lot of dead cells floating.
3- Contamination? If you are worried about contamination, it might be worth adding some antibiotic to your media, just to be sure. 1 % Penstrep should do the trick.
Best of luck! =)
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Tumor suppressor genes (such as Rb, APCPTEN and the ever famous p53) are often downregulated in cancer cells, if not completely lost or dysfunctional.
I'd like to know if there are any tumor suppressor genes that are upregulated in metastatic cancer cell lines/models? If so, could you link to articles/publications/studies of such genes? Are these genes bi-modal? Or have people figured out the molecular mechanism behind these genes?
I'd appreciate any and all help provided.
Thank you.
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Hi ,, Here you can find some example ...
SRPX, PRSS8, PTCH1, and finally P53 are TSG upregulated in cancer and promote cancer survival and spread.
You can easily find article connecting this Up-regulated TSG with invasive cancer.
Ref:
1. Exploiting the p53 Pathway for the Diagnosis and Therapy of Human Cancer.
2. The serine protease prostasin (PRSS8) is a potential biomarker for early detection of ovarian cancer
3. SRPX and HMCN1 regulate cancer‑associated fibroblasts to promote the invasiveness of ovarian carcinoma.
4. Targeting the Multidrug Transporter Ptch1 Potentiates Chemotherapy Efficiency.
Hope this info helps.. My wishes to your research ..
Regards
Suresh
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Many thanks.
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Interesting question
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Hello,
I want to work with Resveratrol against some cancer cells. I want to know your experience about its Non-toxic dose by using MTT, its solubilty and some other things. Please let me know if someone is working with it. Thanks.
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Resveratrol downregulates inflammatory pathway activated by lymphotoxin α (TNF-β) in articular chondrocytes: Comparison with TNF-α. Buhrmann C, Popper B, Aggarwal BB, Shakibaei M. PLoS One. 2017 Nov 2;12(11):e0186993.
Resveratrol Regulates Colorectal Cancer Cell Invasion by Modulation of Focal Adhesion Molecules. Buhrmann C, Shayan P, Goel A, Shakibaei M.
Nutrients. 2017 Sep 27;9(10).
Sirt1 Is Required for Resveratrol-Mediated Chemopreventive Effects in Colorectal Cancer Cells. Buhrmann C, Shayan P, Popper B, Goel A, Shakibaei M.
Nutrients. 2016 Mar 5;8(3):145.
Resveratrol induces chemosensitization to 5-fluorouracil through up-regulation of intercellular junctions, Epithelial-to-mesenchymal transition and apoptosis in colorectal cancer. Buhrmann C, Shayan P, Kraehe P, Popper B, Goel A, Shakibaei M. Biochem Pharmacol. 2015 Nov 1;98(1):51-68.
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Hi everyone
I am planning to use the Qiagen RT2 Profiler PCR array for signaling pathways. Is it really necessary to use the RT2 SYBR Green Mastermixes and the RT2 First Strand Kit with the profiler array? Can I use Applied Biosystems SYBR Green PCR mastermix instead (Cat no. 4309155) as well as my already synthesized cDNAs? The platform is Roche Light Cycler 480.
Looking forward for some help. Many thanks.
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u should use rt2 first strand kit, otherwise, the controls will not work. controls are required for data analsis.
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Dear friends and colleagues,
there is a very important resource on the web that is "Atlas of Genetics and Cytogenetics in Oncology and Haematology". The Atlas is a peer reviewed on-line journal / encyclopedia / database established in 1997 that collects data about cancer genes, solid tumors, leukaemia and other resources. It is in open free access for readers and there are no fees for the authors.
I'm searching volunteers and collaborators that would to write with me reviews about single genes involved in cancer.
I have published several papers so far and I believe that Atlas is a very valuable tool for us researchers, scientists and scholars.
I invite you to visit the Atlas website at http://atlasgeneticsoncology.org/index.html for more information. Here is my latest article published http://atlasgeneticsoncology.org//Genes/GC_EEF1B2.html to show you my working method. Anyone wishing to collaborate with me to write a specific review on a gene can tell me as well.
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I have research and interests in medical genetics on a less laboratory research results level. I rely on the scientific research in Atlas of Genetics and Cytogenetics in Oncology and Haematology nonetheless. Congratulations on your ambitious and important research project! Best regards.
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Which colorectal cell lines that has c-Met overexpression or amplification, dose anyone has any related literature please send me
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H820
MET amplification occurs with or without T790M mutations in EGFR mutant lung tumors with acquired resistance to gefitinib or erlotinib
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recently I am working on a project about the effect of bio-luminescence in cancer research. It would be my honor and pleasure to help me on this way
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First, you have to check whether the transcriptomic data available for the organism you study. based on transcriptomic data you will be able to isolate genes related to luminescent activities by PCR. After cloning with a protein expression vector, you will be able to isolate the proteins.
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Vitamins are useful against various diseases.
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Tretinoin / Retinoic Acid (RA) is a differentiation-inducing agent. Tretinoin for APL drives leukemic promyelocytes to differentiate into mature granulocytes but some of the leukemia-initiating cells (LICs) remain, albeit in lower numbers. Combining RA with arsenic trioxide (AS) leads to better survival of APL patients because AS induces APL cell apoptosis and RA+AS synergize to eradicate LICs via PML-RARα degradation.
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The event of death by metastasis or recurrence is very common, many researchers have linked the tendency of some tumors to re-appear to the presence of occult or dormant cancer cells that may have a phenotype that allows them to remain "hidden" from the immune response. However, the understanding of these cells and the mechanisms that they use to achieve evasion remain mostly unknown. It is critical to understand these cells better and to be able to detect their presence for they are of great therapeutic importance.
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Hi
LGR5 is the best biomarker for detection of cancer stem cells in colorectal patients .
u can browse our my profile and see our paper about that
Regards
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Cancer is very dangerous disease and allopathy drugs are unable to treat. Phyto-constituents of Lady's finger may act against this. 
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CML model is suitable for cancer research?
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TARGET THERAPY AND MONITORING OF CHRONIC MYELOID LEUKEMIA
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There are a lot of published data that showed association between vitD deficiency and many disorders e.g. diabetes, metabolic syndrome, CVD, cancer,...
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Please see the following PDF attachment.
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I am looking for information concerning the effect of publications regarding 5-year survival of diseases, like cancer, on the feelings of the patient. As the statistics are frequently published, and available on many websites with information on serious diseases, I was wondering whether psychological research was done as to the impact these statistics have on patients, dealing with diseases. I have tried to find such research myself, but the overwhelming majority of research I have found deals solely with diagnostic importance of these analysis. I will be most thankful for any advice and references on this issue.
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A high survival rate cultivate hope among patients with specific disease like cancer. If you tell a person that the five year survival is 90%, his hope of being one of the 90 who survived will be high and justifiable. If someone knows that the survival of lung cancer is very low, in most instances they will surrender to their fate
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Hi guys!
Next week I have my first 'serious' interview for a postdoc position job that REALLY interests me. It would be done on Skype, I was asked to prepare 20-minutes powerpoint presentation on my 'previous scientific work'. Since I don't have experience, I am not really sure what does it mean. Should I make a 'classic' ppt with structure: introduction, aim, materials&methods, results, conslusions of all my projects? (Would be difficult in 20mins) Or should it be more like what was the project, why we wanted to study just this, what techniques I used, what I learned, what are my skills for the moment etc?
I will appreciate any advice :)
Thank you,
Marta 
PS: we are talking about cancer research position
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Dear All,
Since I am getting countless messages asking for advice and I am not able anymore to answer to each single one, I will share with you my thoughts - of course it's all based on my personal experience and even if it worked in certain environment, does not mean that it's a guarantee...I think that all depends on the field and your future boss expectations.
But here is a description of how it looked like in my case and few advice:
In my case it was an application for a Postdoc position and in the end I just introduced myself - had a slide about what I studied and where. Then I followed with slides about my thesis project - with typical structure: background/introduction, aim, results and interpretation. I added in that case that the project ended with paper submission at that time. Then I did similar slides about additional 2 project I was working on during my PhD erasmus and also here mentioned the papers. And in the end it worked - I got the position and I am still working there ;)
Meanwhile we had also plenty of phd candidates being interviewed for the position in my lab, some of them also by me, so I can give you also some advice based on this experience.
First of all if you present e.g. your master thesis project it's important to have it structured - with clear introduction and aim. We had few candidates that seemed not to know why they were doing some experiments and this is one of the things that made us reject them. Second, be sure that the presentation is linear and logic - ask someone with more experience for a revision, this really helps :) In the end, since noone expects from you rocket sience data, it's important to show which techniques you learned - our boss e.g. asked people to add the last slide with a list of techniques they learned. But this is up to you, I don't know if this is a common practice. To sum up: you have to sound like PI of your master project.
And be ready for questions like why you have chosen our lab, where do you see yourself in 5 years, read the papers of the group - at least few.
That's it - good luck!
Marta
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several old studies says no, more recent studies says yes. Robert Bell in 1870 used raw salad to increase amygdalin in cancer patients and used saliva to detect the presence. Several sources say amygdaline converts to cyanide in an acidic enviroment and when gets in contact with acid lactic of cancer masses convert to cyanide and kill them (sort of selective killing). To be absorbed as amygdaline and avoid convertion into cyanide in the gut it should be taken from food rather administered directly. There are some roman texts about incresing lifespan of cancer patients with regular intake of 2-3 bitter almonds (rich on amygdaline) a day. Anyone has experience on it? Are research on amygdaline extensive and satisfactory? What is your opinion?
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Role of amygdalin (laetrile or vitamin B17) in cancer treatment is not yet scientifically proven. Please go through the following RG links.
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An exogenous RNA molecule is involved in the pathogenesis of cancer and transfered cancer among animals. It is living organism.
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In addition to viruses, certain kinds of bacteria can cause some cancers. The most prominent example is the link between chronic infection of the wall of the stomach with Helicobacter pylori and gastric cancer.
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Can anyone suggest me the precise and new anti-cancer target for docking of peptide type compounds. As per literature reports the chemical class of the above peptides are mitotic inhibitors. The BAX and BID along with Caspase 3 were highly expressed with peptides while in cytotoxicity studies. When the above genes are over expressing (Biomarkers) is an indication of apoptosis. Can I choose Caspase 3 as druggable target for these compounds and go for docking ?
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If an article suggests the intracellular protein inhibition for the induction apoptosis in cancer would it be feasible to inhibit the pathway by inhibiting the membrane proteins or receptor tyrosine kinase, as the peptide based drugs are more of surface acting molecules
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Cancer research has developed precise biomarkers to use a proper cure for different kind of cancers. Why there is no proper biomarker to find the precise treatment in autoimmune desease, such as psoriatic arthritis?
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I Know. Thanks anyway for aswering me.
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Since humanized and SCID mice are very expensive to procure and maintain. I was wondering to develop a cost and effective approach to develop an in vivo model for cancer research. Thymectomy is well established method to cause immunosuppression. I was wondering what will happen if we try to develop PDX model in thymectomized alone or in combination with immunosuppressants in C57bl/6 or Balb/c mice. Will tumor model be developed ?
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I DONT KONW
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I woul like to submit a Review article in the Journal of Cancer Research and Clinical Oncology. Are there publication fees for this Journal?
Thank you in advance!!
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Thank you to everybody! Is it the same for Current Hematologic Malignancy Reports Journal?No publication fees?
Thank you
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Hello everyone,
I have just started using Olympus fluoview for confocal microscopy and I don't know much about microscopy. I plate my cells (HeLa) in 4 welled Chambers slide. Stain the cells with Mito tracker dye, fix it with 4% PFA, wash, add mounting media and do the microscopy. The first time everything was fine. The second time I was trying to optimize the dye concentration, but something went wrong. The microscope kept on showing 'The focus position not found in scanned containers'. I had two slides and both of them had the same problem. When I tried with my old slide, it was fine. I have no clue what is wrong with my slide. All the chemicals and reagents that I have used is same as before except for PFA. The second time I diluted 8% PFA with sodium phosphate buffer to get 4% PFA. Can PFA cause problem finding focus?
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Apparently I went back to my old protocol and it worked fine. I don't remember the detail since it has been a while.
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We are trying to test some formulations of PD1 antibody on cancer models and would like to have a robust in vitro technique for future in vivo assessments.
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If you want to measure the effect on cytotoxicity you can just use it in a standard coincubation cytotoxicity assay using overnight activated PBMCs for example.
You can then compare killing rates between treated and untreated samples.
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Most studies on CAR-T cells use NSG mice (B/T cell deficient) based on the papers I found. Only a few use nude mice (T cell deficient, B cell present), which is what we are hoping to do. Does anyone have experience with using nude mice tumor model in CAR-T cell studies? What are the problems with using nude mice for CAR-T cell studies (since B cell is APC), and what are the potential solutions? Thanks a lot for any responses!  
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Hi Shirley,
Thanks for your rely^^.
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In my job, I transient transfected Active-wnt3a construct into Hek293T and Hek293 for expression , I collected three batch(fresh medium 5ml in 10cm dish for 24hr)
conditioned medium and concentrated in 500 µl ,but it didnt have any effect when it treatment 20µl for TopFlash repoter assay!!!!
Can any one tell me what can i do?
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In support of Svitlana’s answer, i can add my personal observations. I did a secretion assay on 293T cells expressing Wnt3a and found barely detectable Wnt even after concentrating the conditioned medium 30 times. Wnt3a was robustly expressed (as evidenced by western blot on whole cell lysate) but curiously i could not detect the higher mass (glycosylated) form of Wnt3a. Svitlana, do you know if Wnt1 is secreted by 293T cells?
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I have submitted a manuscript to clinical cancer research and the status changed to "decision pending" on the same day "under review". And I wonder whether it is not good news, how long will I receive the email from the editor?
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It is anybody's guess. It is likely that the journal has disclosed its review procedure policy at its website. If the preliminary review by the editor is a prerequisite for review by reviewers, it is likely that a "decision" to review the manuscript or not to review is "pending" . Therefore, it can be interpreted both ways.
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We are looking for highly motivated candidates to join our lab. More info available in the link below:
Any help with spreading the word about them will be highly appreciated.
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Thanks for sharing this info.
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Hello! For my study assignment (cancer research, up/downregulation of a gene in high and low density cells) the standard deviation is not necessary but we think it is important to put in our short communication. We found out that the standard deviation is taken from the delta-CT value. For example: you have a gene of interest and a reference gene. Each gene is done in triplo and you have a control and a treatment.
GOI control: 32.37, 31.76, 31.80
Ref control: 16.05, 15.10, 15.76
GOI treatment: 30.1, 29.78, 29.88
Ref control: 18.56, 15.64, 14.76
We've calculated the delta-CT from the averages of the measured values. How can I calculate the standard deviation for the delta-CT value?
Thanks in advance for your help.
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The data you show is for a single dCt value (even if this is determined as an average from several technical replicates) from a single biological sample (actually two: one "control" and one "treatment" [assuming the last line is a typo and should read "Ref treatment"]).
If this is all you have, then there is no way to estimate the SD between samples. The SD between technical replicates is usually not interesting (exception: if you want to assess the quality/reliability/reproducibility of an assay - what does not seem to be your question).
If you have a group of more than one biological sample, you can calculate the SD of the dCt values for these samples as usual.
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I'm a Research associate at 57357 hospital , Egypt , I would like to ask about two cell lines : RD , RH30 ( Rahbdomyosarcoma ) we couldn't find them in Egypt and we are committed to certain budget , deadlines and masters thesis based on this project , anyone know a trusted source  or a lab in any country usually uses them to give as aliquot ? Thank you in advance !
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What could we change in preparing tissue samples prior cryosections(cryostat) method for more convenience and results? ( I mean cryo-embedding) The second question is in which areas of research do you use this one and why is it a pretty rare method of imaging? It is interesting to get answers from researchers who are working currently with this method or those who worked with that recently.
Thank All in advance for any ideas.
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Hi Dimitri;
As our lab does immunochemistry and histoenzymatic I can tell you by personal experience that frozen sectioning improves results in many ways; as the tissues are not fixed, epitope retrieval steps are not needed in much cases. And the histoarquitecture is pretty well conserved, endogen eznymatic activity is also conserved.
hope it helped!
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I'm starting to work on microRNA profile in a mouse model of liver cancer. We would like to use the next generation of sequencing approaches since it enables mapping and quantification of all transcripts.
Since it is the first time we are working with sequencing, I was wondering if somebody could suggest the best company to contact. I was in contact with LC Science because a colleague of mine recommended them. We are also evaluating the Arraystar sequencing service. Does anyone have some suggestions or recommendations for other companies?
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I m also interested to do miRNA sequencing in exosomes.
Pl inform the good company in India and how we can submit the samples for analysis?
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To fight the disease effectively, researchers from across the scientific spectrum and beyond must join forces.
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Yes
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what is the effect of aloe vera on cancer cure , Is there any approved benefits from using aloe vera with chemotherapy in cancer 
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Hi everyone,
I plan to perform CAncer Personalized Profiling by deep Sequencing (CAPP-Seq) for a cancer research project.
Is there any available genome analysis company that uses Illumina Hiseq or a similar NGS system and that can perform CAPP-Seq with a reasonable price?
We will need logistic support for cargo shipment from İzmir in Turkey as well.
Best,
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Can Kucuk Mail gonderirseniz yardimci olacaklarini dusunuyorum.
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Is there any experimental data about structural implications for the mechanism of action and cellular specificity of both Cry and β-PFT type of parasporins?
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Hi everyone,
I'm looking to find a way to determine the concentration of the ketone body Beta-hydroxybutyrate (BHB) within cell culture media. Preferably, a method that would employ a spectrophotometer. Is it possible to make up a solution of known concentration and run it through a variety of wavelengths in order to determine the lambda max for the ketone body? Once I have the lambda max, shouldn't I be able to determine the concentration of BHB within a solution by performing a serial dilution of BHB solutions of known concentration, then comparing the unknown solution's absorption to those of the standards? 
If anyone has a better way to determine the concentration of BHB within a solution, then I would love to hear it. 
Thanks in advance!
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In urine I used as screening test based on the principle of Legal`s in an modified micromethod described below.
Reagents
Alkaline solution: dissolve 1.2 g (3 M) NaOH in 10 ml dH2O. Store in opaque plastic bottle at room temperature.
SNP stabilized reagent: dissolve in 5 ml dH2O, 0.25 g sodium nitroprusside dehydrate and 0.175 g sorbitol. Store in amber glass bottle, protected by light, resulting stable at room temperature for 1 year.
Procedure
Dispense in MTP well 200 µl of urine, add 40 µl of alkaline solution, mix and then add 20 µl of SNP reagent. After shaking the mixture, a red color appears, caused by creatinine, and add 40 µl of glacial acetic acid that in the presence of ketone bodies the red color turns to red-violet.
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Transhiatal esophagectomy was championed by American Dr. Orringer since 1976. The issues were: inadequate field exposure; inadequacy of node clearance, use of blind & blunt dissection & (rarely) azygos vein tear. Benefits: avoiding thoracotomy and all the morbidity associated with it.
So, what is the current opinion?
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....Advantage
My only comment on squamous cancers is that because of their high propensity for nodal spread up the mediastinal chain that this makes them relatively unsuitable for TH orsophagectomy other than the fact that if combined with neo adjuvant chemo or chemorad that the disease may already have responded. We do have a series of squamous cancers with very acceptable outcomes after THO (45% 5YS)
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Hi
I was wondering if anybody have used stain free gel (Biorad) to compare the expression of a protein of interest in Cytoplasm and Nucleus?
If I were to submit a paper including this stain free blot image to a decent journal, say Cancer Research, would that be any issue?
I am wondering whether I have to go for conventional normalization method(using loading control) for comparing proteins from different compartments in cells. While stain-free blot is easier to work with, I am worrying that reviewer would still like to see loading controls of nucleus and cytoplasm.
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Hi Bumjun,
You are welcom.
The positive control, to prove your antibody is working. Whilst, the striping procedure is to prove the availabilties of your protien samples (if you get negative result).
Feel free for any further qeustion, please
Best wishes
Sarmad
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Hello fellow Patchers and Sealers,
                                I made a pipette solution recently and expected the osmolarity to be around 270 mOsm approximately according to rough calculation but when I measured it it was only about 191 mOsm. I am pretty confident I did not miss out on any ingredients or added more water that could decrease the osmolarity than expected. Now, at this point, I am requestioning my manual calculation of 270 mOsm (approx) from the recipe. It would be really helpful if someone can proof-check my calculation for the recipe below. Just the K- gluconate itself should make it 244 mOsm to begin with.
NOTE: To adjust the pH to 7.4 I had to add about 1 mL of 1M KOH. I wonder if this could be a reason. I doubt it though since only an mL of this in a 100 ML final volume should not make the difference.
Recipe: K-Gluconate 122mM, NaCl 9mM, MgCl2 1.8 mM, Mg-ATP 4mM, Na-GTP 0.3 mM, Creatine phosphate 14mM, EGTA 0.45 mM, CaCl2 0.1 mM, HEPES 10 mM
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Dear Maibam,
gluconic acid is weak so its dissociation is incomplete in water. You should increase the amount of K-Gluconate or another component of your solution in order to reach the desired osmolarity.
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Does anyone have a running model of a breast or ovary tumor?
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What is the different of B16 & B16-F10 melanoma cells? And which of them can be use for subcutaneous tumor models in C57BL/6J mice?
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