- Prem Subramaniam added an answer:3Why would a serum sample not freeze for more than a week at zero? Tiger top properly drawn, settled, spun, and serum transferred to plastic t tube.
This was drawn for an ENOX2 test. The patient had mildly elevated Calcium, TP, Albumin, and ALT. All other labs were normal except a chronically high Lp(a).
Patient developed Shingles at about the same time.
In case of less comlex solutions, sometimes you can get a supercooled state,. In this case, if you just lightly shake the tube it will freeze right in your hand ! In case of FBS, I would add that you should make sure your freezer is at -20C to get it to freeze. The door of these freezers is much warmer than the inside, especially if you keep opening the freezer often, and even PBS solutions sometimes do not freeze. In the latter case, then you know it is not keeping temperature.Following
- Kambiz Gilany added an answer:6I am interested in finding the markers for urinary bladder cancer for diagnosis. Please help me in finding the markers via microarray.Working plate form.
Maybe it is to late but I think you can look at the following paper Microarray- and metabolomics study:
PMID: 20059769 Microarray study
PMID: 24721970 Metabolomics Study
I hope it is useful...Following
- Go J Yoshida added an answer:1What is the most efficient method for isolation and sorting myeloma stem cells?
myeloma stem cell
Given that multiple myeloma arises due to the monoclonal expansion of abnormal plasma cells, which leads to the production of monoclonal antibody characterized by Bence-Jones protein, IgG, IgA, IgM, which is produced in a mutually exclusive manner depending on a clinical case. That is why the typical concept of cancer stem cells in the heterogeneous tumor cell society is difficult to find in multiple myeloma.Following
- Mario Valentino added an answer:13Does anyone know distinguishable markers for arterioles, capillaries and venues for doing immunohistochemical staining?I am looking for specific markers for arterioles, capillaries and venues for doing immunohistochemial staining by which I can distinguish them properly when I visualize the section under the microscope.Following
- Roman Mezencev added an answer:3How to calculate the detection limit from the calibration curve?
Hello Every body,
I am working on detection of cancer biomarker and I get the calibration curve.
I calculate the detection limit by obtain the 3*std/slope. but the result doesnt make any sense !
I used range of concentration and the response is current. I got the std of the current values and multiply by 3 then I substitute the result in the calibration equation to get the concentration but I got value higher than my smallest value of concentration that used to generate the calibration curve .
You are welcome Mohammed. In the link below you can find a very nice and concise discussion of both methods that I mentioned above.
- Ijaz S. Jamall added an answer:99+Cancer a mere matter of luck? Or is there something under-appreciated?
It has been all over in the news lately: The majority of cancer is obtained by bad luck, not by lifestyle or inheritance. See attached.
Really? The data appear solid and they make sense, but the conclusion seems a bit premature: the observations are based on established risk factors in the USA and I assume (let the experts please come forward!) that these risk factors are based on occurrence. This means we do not see all those cases where the patient's immune system adequately takes care of the anomaly.
How does occurrence of cancer relate to failure of the patient's immune system, and can we monitor this based on adequate biomarkers? How will the statistics and the conclusions change when this factor is included in the analysis?
@Giovanni- makes complete sense. And, to add to your excellent points, the metabolic components are highly variable and subject to the biochemical demands of the particular tissue at the time-so there is a substrate variable and a time variable that must be considered.Following
- Mahendra Kumar Trivedi added an answer:5Are there some complete and authoritative databases for cancer biomarkers?
As we know, it's a time-wasting work to curate biomarkers of some specific cancers from massive literature and they may be incomplete for us. Therefore, I want to know whether there are some complete and authoritative databases for cancer biomarkers? By the way, noncommercial and publicly available databases may be better.
You can take a reference with my research, on cancer biomarkers
Here I am sharing my research paper link to you.
I hope help you .
Mahendra Kumar TrivediFollowing
- Swathi Reddy Pullagurla added an answer:12Can anyone recommend instrumentation for quantitation of exosomes?I tried out a Nanosight instrument which I like very much but is very costly.
Could you please forward me information on Guava flow cytometer for exosome analysis. My email is email@example.com
- Marianne Levon Shahsuvaryan added an answer:18Do you think biomarkers are the future of medicine?Http://acceleratingscience.com/proteomics/strategy-for-cancer-biomarker-verification-using-srmmrm-mass-spectrometry/
A multicenter study led by Christian Haass and Michael Ewers of Ludwig-Maximilians-Universitaet in Munich has identified a biomarker associated with the activation of an innate immune response to neural damage during early stages of Alzheimer's disease.Following
- Kaleem Ullah added an answer:8What are the possible ways to prove optically that this cell is effected from cancer?
Hello everyone, i have oral skin sample which is effected by a cancer. I have no idea about what kind of cancer is and what looks like a cancereous cell because i have no biological background. So any biophysicist or biologist would like to help me. In my last question i have attached its picture now i am attaching it 2nd time.
@Chiara Mignogna Thanks for your reply. It is on of the cell of the skin oral sample on a glass slide. I have measured it by IT two-photon microscopy. Please have a look at one more image:Following
- 7How can I treat cells for detecting autophagy marker (LC3 and p62) perfectly?
Recently, I tried to measure autophagy marker LC3 and p62. I starved MEFs cells with EBSS or DMEM (no glucose, no glutamine), but the problem was I could see strong signaling of LC3-II in case of untreatment. Moreover I also checked the LC3-II in K562 cells (leukemic cells) and got the same result. Why do I always see strong LC3-II band in untreated cells? I also tried the Beclin 1 KD MEFs, no difference of LC3-II between parental MEFs and Beclin 1 KD MEFs after different time of starvation (1h, 2h, 3h, 4h).
With all the experiments I tried, the problem is I always see strong LC3-II signaling in negative control group (untreatment or Beclin 1 KD cells). How can I design experiments to avoid seeing LC3-II in untreated cells. The LC3 antibodes I tried are from CST and novus bio. In the case of K562 cells, please see the attached figure. Thanks
The attached article could be of help.
- 7Can anyone suggest a reliable glioma marker?
We're looking for a nice marker to detect tumor cells within glioma tissue. The tissue will be dissociated into a cell suspension. After that, we want to stain the suspension with a mixture of antibody-conjugates for flow-cytometric analysis to get an idea of the cellular composition of the tumor...
Can anyone suggest a marker (either extra- or intracellular) that is only/mostly expressed by tumor cells?
Galectin-1 could be interesting (see the attached articles).
- A.Baki Agbas asked a question:OpenI have made a calibration Curve from pooled plasma. Is this dose/response curve legitimate?
I am working on a development of blood-based biomarker protein for AD and ALS; however, protein candidate is not readily available in purified form. Therefore, I have the problem of not having calibration curve to quantitate this protein. I was thinking that if I collect , say 50 plasma samples and I know that the bio-marker protein is elevated. Pooled these plasma and construct a calibration curve based on their total protein content. Now, I have a kind of dose/response curve. I am asking my fellow researchers that is this way a legit. I know it is not ideal, but i stacked with the absence of the purified form of this protein. We have attempted, but it is quite unstable. Any recommendation, thoughts, tips are welcome.
- Sandeep Telkar added an answer:4How to construct TGF-β induced EMT( Epithelial to mesenchymal transition) pathway?
In my work am trying to construction EMT( Epithelial to mesenchymal transition) pathway induced by TGF-β but I am finding difficulties while construction i.e.,
whether pathway constructed for particular cancer is effective or in general
(because the TGF-β downstream signals are same i e SMAD and non-SMAD signal are same for all type of cancer)
thank you Amira Ouanouki,Following
- Horatiu Cristian Popescu added an answer:2Why mutant p53 expression is increased in colorectal adenocarcinoma obese- type2 diabetes mellitus patients?
Is there a particular feature or a direct connection that can explain why immunohistochemical expression of mutant protein p53 is increased in obese and diabetics (type 2 diabetic) patients at diagnosis of colorectal adenocarcinoma compared to non -obese diabetics versus non-diabetics obese/non obese?
Study included immunohistochemical analysis of 2 proteins Bcl2 and p53 (both monoexpression and coexpression) in 100 newly diagnosed colorectal adenocarcinoma specimens divided equal in 2 groups diabetics (type 2 diabetes mellitus already existing at diagnosis of colorectal cancer) and non-diabetics .The preliminary statistics results have not indicated a significant difference between the 2 groups, in terms of Bcl2, but indicates an increased expression of p53 only in the diabetic-obese group compared to non-diabetics obese or non-obese (statistically significant ) which theoretically indicates a contact point between obesity, cancer and diabetes. Interestingly moderately low degree of differentiation is associated with both diabetes and the presence of p53 positivity; G2-p53 associations is an intriguing part. Bcl2 expression was low and insignificant in both group.
Thank you for your clear and concise information you gave me that will help me a lot. Best, HoratiuFollowing
- 2Most of the biomarkers of GBM based on proteins are common to other cancer also (EGFR,VGEF,PTEN,MGMT etc). How can it be specific for glioma?What is the most specific marker of glioblastoma. Where this biomarker science leaning toward?
I fully agrees with Reza's responses / comments.
You must clearly "subdivide" the terms biomarkers in:
- diagnosis (GFAP is a specific marker of astroglioma, but can also be overexpressed in reactive astrocytes)
- treatment responses,
the attached articles could may be of help for you.
- Héctor Osorio-Vega added an answer:5At this moment, are there any specific markers to detect the presence of cancer stem cells or cancer dormant cells in soft tissue?
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.
currently my interest is in Colorectal cancer, this one have a very predictable behaviour, however, i haven't been able to find any specific marker. The best option is to target the overxpression of MMP-7 and/or beta Catenin, but i would expect them to not be so present in residual cells that are dormant, or in cancer stem cells that have a very slow metabolism.Following
- Alejandro Martin added an answer:2Clone having repetitive sequence insert and its discripancy upon scale up?
I have cloned my 3.3Kb insert having repetitive sequence in yeast expression vector, by directional cloning in E.coli. Positive Clone of 8.7Kb was confirmed by restriction analysis. While the same positive E.coli transformants when grown in 100ml LB, the restriction pattern observed is different. But the the digestion pattern is as expected when it is isolated from 5ml culture. Could anyone please let me know why this discrepancy observed from 5ml to 100ml scale up.
The more doublings of your population, the higher the probability that mutants bearing deletions will arise and overtake the culture. Most likely you already have mutants in the 5 mL culture, but at proportions low enough to pass below the limit of detection of your assay (restriction digestion + gel electrophoresis).
Have you tried E. coli strains designed to propagate repetitive sequences, such as SURE or STBL3?Following
- Massoud Vosough added an answer:3What is marker of malignant transformation in cultured cells?
I am planning to culture MCF10 cells with different carcinogens,can some suggest a marker which I can look for as a sign of early malignant transformation.
The epithelial-mesenchymal transition (EMT) is a process in which epithelial cells lose their cell polarity and cell-cell adhesion, and gain migratory and invasive properties to become mesenchymal stem cells. E-cadherin is a protein that in humans is encoded by the CDH1 gene and down regulated during early stages of carcinogenics.
- Volker Gatterdam added an answer:3Where we can order cancer aptamers?
Sensing with the help of aptamers.
maybe you can also find interesting "aptamers" at SOMAlogic (http://www.somalogic.com/). They are selling small binders, which are a hybride between DNA and Protein.
- Stephane Chartier added an answer:1What should be positive control for IHC and western blotting during experiments to prove whether an untested protein could be a biomarker for cancer?
I'm going to design a study to test whether an untested protein could be a biomarker for pacreatic cancer. I intend to do immunohistochemistry of the pancreatectomy tissue of the patients with pancreatic cancer, benign tumour and disease - free. I will also do Western blotting and ELISA. Could you please suggest me in terms of positive control selection for these tests?
Hi Saif! I'm not too familiar with Westerns and ELISA but I have done extensive IHC work on muscle and skeletal tissue in mouse cancer models. Typically you'd check the data sheet on any given Ab to see what positive controls the manufacturer recommends. Given that your Ab is untested, I would simultaneously stain for an Ab which will be widely expressed in any tumor alongside your untested Ab. I would recommend CD68, a marker of the myeloid lineage and macrophages. Also, CD31, a marker of endothelial cells is expressed robustly in tumors. Hope this helps!Following
- Vladimir S Kurćubić added an answer:6Do you know what is the level of MMP-9 in the breast cancer tissue?
Do you know what is the level of MMP-9 in the breast cancer tissue?
Nema na čemu.
- Saif U. Sikdar added an answer:5Is it possible to validate an untested protein as a biomarker for pancreatic cancer solely by immunohistochemistry?
I want to design a study to assess whether an untested protein could be used as a biomarker for human pancreatic cancer. I intend to do immunohistochemistry of this protein in pancreatic tumour tissue and normal tissue. I am thinking of doing TMA and normal tissue microarray. Should I include any other assays like ELISA of body fluids and Western blotting for validating this protein?
I have also found people doing MALDI-TOF and MS. Which test will be more practical for clinical diagnosis?Following
- Graham Dellaire added an answer:3What are the main biomarkers/ molecules to study in HNSCC in DNA damage pathway because of radiotherapy?
We are following HR and NHEJ pathway. Want to know the assays and techniques to go ahead
Re: 8-OXOdG ... Not sure how useful (or appropriate) detection of 8-OXOdG is going to be for general surveillance of DNA double-strand break (DSB) repair by HR and NHEJ.
8-OXOdG is a measure of base-excision damage and the BER pathway has nothing to do with NHEJ or HR, which are DNA double-strand break repair pathways. For general DSB repair competence, good old phospho-S139 H2A.X (i.e. gamma-H2A.X) is the gold standard for most IR dose experiments especially in vitro. If you cells are cycling, BRCA1 or RAD51 foci formation are good markers of HR ability. Otherwise you need to do reporter assays for NHEJ and HR specifically.
We developed one such assay for HR, that relies on CRISPR-mediate homology directed repair, see: Pinder et al., 2015 NAR
Cheap, quick, fast.
- Govind Babu K added an answer:2I am creating a roadmap for high risk wilms tumor for a research institute. Does anyone have a cell line data for high risk wilms?
These cell lines along with available xenografts and other factors along with other biomarkers ( if you have any it would be appreciated) will be used to build a preclinical model for high risk Wilms tumors. The development of assays specific to this model will result in the ability to do novel cell line and animal testing which could potentially lead to clinical trials with new compounds for children with these disease.
- Yaser Arjeyni added an answer:4What are the transcript variants for BRCA1 and where can I find the BRCA1 exons?
I know BRCA1 on ch17q21 have 29 transcript variants I found 7 of them on NCBI but, I couldn't find others.
Can anyone please help me find them?
Also I know BRCA1 has 24 exons, but where can I find their range on DNA?
I would appreciate any help.
You have not specified aim of your study but overall i think the variants you have found on NCBI must be responsive for your survey.Following
- David S Guttery added an answer:1Is it possible to use CAST PCR technology for cell free DNA analysis?I am trying to find out the most "sensitive" methods for detection of specific DNA mutation in serum/plasma samples. Does anybody have any knowledge on the issue?Following
- Samah Oda added an answer:14What is the role of LC50/LD50 value of any compound or metal for toxicological or other studies?Why do we calculate only LC50? Why not 30, 40 or other values?
How could I calculate the acute toxic dose from LD50? Is LD50 is acut doseFollowing
- Luboshits Galia added an answer:3How can I find the complete profile of specific marker production by individual cells?
For example what proteins and receptors are produced in normal breast cells and malignant cells.
you can run protein antibody array and compare the expression in normal versus malignant tissueFollowing
- Lin-Lin Bu added an answer:6NF-kB and JAK/STAT3 pathways crosstalk, (IL-6??)?
I am looking at studying the connection between NF-kB and JAK/STAT3 in normal and cancer cells. I am pretty certain from my research that there is a connection with IL-6. What type of tests should I propose to test this and what would some expected results be?
“NF-κB and STAT3 – key players in liver inflammation and cancer” maybe this review could have some help for you.Following
About Cancer Biomarkers
A biological molecule found in blood, other body fluids, or tissues that is a sign of a normal or abnormal process, or of a condition or disease. A biomarker may be used to see how well the body responds to a treatment for a disease or condition. Also called molecular marker and signature molecule. A cancer biomarker pertains to any biomarker that fits the aforementioned definition but only for cancer, and no other disease.