Liver - Science topic

I have a problem concerning my liver fluorescence staining: one primary antibody, GARP, in the attached picture in red, is causing these weird stainings on and around the nucleus. Does anybody happen to know how to improve my staining and get rid of these false positive stains?

Or general advice for liver staining, as the autofluorescence in liver tissue is always really high.

We are cooking our samples in citrate ph6 buffer, and using Avidin+Biotin as well as Tris,TWEEN20 + 5% BSA for blocking. In order to attach the fluorescence dye we are using Dylight 550.

I would very much appreciate any advice!

The US CDC knew very early, from post-marketing Adverse Reaction reports, that the COVID-19 vaccines were causing numerous Amyloid fibril diseases including Amyloidosis subcategories: Cerebral angiopathy, Senile, Cardiac, Cutaneous, Dialysis, Hepatic, Pulmonary and Renal.

Elegant work by Swedish researchers demonstrated that Synthetic Spike Protein can be broken down in vitro by incubation with the protease Neutrophil Elastase into short peptide sequences that can induce damaging amyloid-like fibrils.

References: Vaccine Adverse Event Reporting System (VAERS) Standard Operating Procedures for COVID-19 (as of 29 January 2021)

Coronavirus disease (COVID-19) pandemic is a global health problem. Infected patients usually have respiratory symptoms due to lung involvements. However, liver impairments could be another findings.

So does Covid-19 affect liver functions & how?

Hello everyone,

can somebody recommend me an fgf21 Elisa kit which actually works on tissues such as skeletal muscle or liver?

I ve found a lot of kits that has tested on serum with good results, but I am not sure about their efficiency on tissues.

Thank you in advance.

In partial hepatectomy models, fat accumulation occurs during hepatocyte regeneration. Whether it is needed is controversial. Though it was initially thought to be the "fuel", later experiments have shown that proliferation can proceed even if fat accumulation is disrupted. My question is, what triggers this accumulation and what is its function.

Thank you

Literature suggesting hepatitis / liver injury following COVID vaccination. A latest peer reviewed article suggests SARS- CoV -2 vaccination (Pfizer)can elicit CD8 T-cell dominant Hepatitis (Autoimmune hepatitis).Details attached in document .Should such rare side effect be taken into consideration during vaccination?

I am going to use healthy liver cell line for the cytotoxicity experiments with nanomaterials. So, I am wondering that are there any differences between THLE-2 and THLE-3 cell line ?

Hello !!!

I am working on the hepatoprotective effect of some medicinal plants and, i want to use the ccl4 induced acute liver injury ( 7days protocole)

there are a plenty of articles that treat the subject, and each one use differente concentration !!

so according to your experience what is the best and most used ccl4 concentration to induce mice acute liver injury ??

We made a pilot study with N=2 mice per group to test biodistribution of rhodamine 6G-loaded silica nanoparticles functionalized with PEG, at 1.5h, 6h and 24h. We used GFP detector in IVIS Spectrum 200 imaging system and normalized fluorescence efficiency to control organs average, as a ratio (N=2 as well).

We expected to find high accumulation in liver, spleen and lung, possibly decreasing with time, but surprisingly we cannot see any rhodamine signal in liver at any time when compared to control organ. We made sure the biliary vesicle was not present and liver was properly washed. Signal in spleen is not very high either. Nevertheless, lung signal at 1.5 hours is 5-fold the controls signal, decreasing in time and disappearing at 24h.

We know optical imaging is not the most sensitive technique to assess nanoparticle biodistribution, but we can clearly see rhodamine signal in lung. What could possibly be going on in liver and spleen? Could their color or opacity cause any quenching of the signal? I've found this article detecting rhodamine in liver using the same equipment as we do () so I guess it is not a matter of quenching...

Do we have the best and most biocompatible nanoparticles ever? I don't think so! But we need an explanation for the lack of liver uptake...

I have been performing immunofluorescence staining for liver tissue that is embedded in paraffin. I have been getting inconsistent results and the full liver section never stains through properly. There are always parts of the section that don't stain at all, or stain very poorly.

I know that the antibodies work well. I'm not sure if there's an issue with paraffin removal or antigen retrieval? I heat the sections for at least 30 min at 60C before the xylene step. Then I do the following:

[2X] Xylene, 7 min

[2X] 100% EtOH, 5 min

[2X] 70% EtOH, 5 min

[3X] rinse in autoclaved water

Antigen retrieval in 1x DAKO Target Retrieval reagent in pressure cooker.

I am attaching an example picture. I would greatly appreciate any trouble shooting help!!

The characterization of heavy metals like gadolinium in organs like liver, brain, lungs can best and simply be done by.....................

Dear all,

Which is the best mounting medium for liver frozen sections stained with oil red O? An aqueous one as Ab104135 could be the solution (even if suggested for IF)? Thanks in advance.

I did a qpcr reaction with SYBR master mix, and cDNA from epididymal adipose tissue. The RNA used for cDNA synthesis was intact (two bands on the electrofluoresis gel), good A260/280 and A260/230 ratios. With the cDNA from adipose tissue it did not amplify none primer. I did it with a liver cDNA and it worked. The tested primers were from genes such as PRDM16, UCP1, Beta Actin. Not even with the one of Beta actin it amplified. Could it be some inhibitor present in RNA or cDNA? Any suggestion?

We have realized a toxicological experiment with Wistar rats. Liver was collected and fixed wtih formallin. Histopathological analysis was performed from HE stained sections. Now, I am looking for a more specific method for analysing liver lesions from these formallin-fixed material or embedded tissue. Unfortunately, I do not have molecular techniques nor immunohistochemistry. Is there an histochemical method or histological staining specific necrosis detection? Thanks for helping!

While a pt. Use HMG-CoA reductase inhibitors , it cause increase in liver enzyme ( 3 times the normal) and stop HMG-CoA reductase inhibitors. then it expect that liver enzyme will go down & when it become normal can we reintroduce statin again?. And when to recheack liver function again? I mean after how many wk. Of use of statin recheck liver enzyme again?

I have two chemotherapy drug (A and B), at baseline, after first cycle, second cycle and third cycle, I want to compare between two drugs after each cycle and and which drug associated with more adverse events with time at the end of third cycle and after each cycle?

Some safety data is continuous such as liver enzyme, and some is categorical such as grade of some adverse events.

I think for continuous data repeated measure ANOVA can help but what about categorical variables?

Hi all,

I have been working on optimizing lysis conditions to do whole proteome lysis from liver tissue and have a head-scratcher. Using the BioRad detergent compatible BCA analysis kit, I get a woefully low estimation of protein extracted when compared to doing a mass balance (weighing empty tube, liver, then remaining pellet after extraction)... I've washed the liver tissue as much as possible to remove blood (non-perfused at harvest) and the samples aren't bloody looking. Does anyone have any suggestions of expected protein extracted per wet liver weight? If I know that, I can at least have a better idea of which number I should use (BCA or mass balance).

many thanks

protocol or any relevant article please share

Hi,

I am starting to work on a liver histopathology image classification project and looking for publicly available datasets. Any guidance related to this will be really helpful for me.

Plaques form due to a self-healing mechanism of blood vessels and will increase over time. When entering blood vessels, they block blood flow, lead to hypertension and decrease blood flow to organs such as the heart. To get rid of these plaques, we need to boost the good cholesterol such as HDL or improve health of liver to produce enzymes that move these plaques. So, what other ways to get rid of these plaques without using invasive methods?

Thanks and best regards.

Currently I'm trying to adjust Oil Red O staining method for liver lipids. Base protocol is A. Mehlem's (Nature Protocols; Vol. 8, No. 6; 2013; p. 1149-1154). I tried different times of ORO staining and counterstaining with hematoxylin. Had some problems with non-specific staining of cracks between cells which might have been solved with varying times. As I believe in the pics the darker dots are lipids and red colour between cells is other type of lipids? Maybe someone who has experience with ORO staining method could explain if attached pictures looks good and if not maybe suggest some corrections which might improve staining?

Liver samples are from chickens treated with high concentration of TBT.

Sample 7: 5 min. in ORO, 15 s. HTX, 30 min. wash; S8: 5 min. ORO, 1 m. HTX, 60 min. wash; S12: 5 min. ORO, 3 min. HTX, 30 min wash; S13: 5 min. ORO, 1 min. HTX, 30 min. wash.

I have two unique teleost species, Saffron Cod and Broad Whitefish. I am analyzing the HSP70 concentrations in the cranial, hepatic, and muscle tissues. The Broad Whitefish samples all worked perfectly. However, the liver and muscle tissue samples for the Saffron Cod aren’t running (the cranial tissue samples are working perfectly). The HSP70 protein seems to be aggregating and sitting on top of the well.

My advisor and I believe perhaps the cod have a higher concentration of fats and thus aren’t separating like the other samples. All of the tissue samples were prepared the same way using a homogenization buffer with 4% SDS and Tris-HCl. The tissue samples were homogenized in the buffer, boiled for 5 minutes and spun down at max RPM for 15 minutes before extracting the supernatant with the protein sample. The protein samples are placed in 2xSB and boiled for 3 minutes before being loaded in 10% gel. All of the solutions, both cod and whitefish, are clear and fluid when at room temperature (none are gelatinous or are behaving weirdly). I have attached images of the gel and Western Blot with four Broad Whitefish samples (BDWF) and four Saffron Cod samples (SFCD). Muscle tissue is MT, hepatic tissue is HT, and cranial tissue is CT. The Western Blot for the Saffron Cod muscle sample didn’t show up for some reason, but when I ran it the other day it looked exactly like the liver tissue; the band was sitting at the very top of the well. Thank you for any and all help!

Any idea how long cholangiocytes live?

Average life span of cholangiocytes?

Can we combine alcoholic and non-alcoholic liver disease into a single fatty liver risk scoring system?

If no, why?

If yes, what will be the potential of such research and implications.?

Hello,

I'm trying to find suitable antibodies to discriminate hepatocytes from other cell types in the mouse fetal liver. I was considering ASGR1, but I found that it can be expressed in monocytes as well, or ABCC2/MRP2, but I couldn't find a nice antibody (please help in case you know one).

Any good (fetal) hepatocytes marker would be good for me.

In an ideal world, I would prefer a conjugated antibody against a surface marker for flow cytometry of live cells, but also an unconjugated antibody for immunofluorescence (in this case it can be an intracellular marker). I want to confidently distinguish hepatocytes from other cell types such as endothelial and haematopoietic cells.

Many thanks!

Investigation of apoptosis in liver tissue

Labeled Liver Tumor Dataset Images

I am conducting experiments to repair the meniscus injury of rabbits by transplanting cells, and rabbits are also given daily injections of tacrolimus, an immunosuppressant.Now I want to test whether the transplanted cells have adverse effects on the liver and kidney functions of rabbits, but I don't know which liver and kidney items need to be tested.I hope friends who have had similar experience can give some suggestions.

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with augmented malaria infections. Thanks. Patrick. I need antibodies that react to mice hepatocytes. And also mouse cd8 and Tcells.

This study was carried out in order to find out the level of sheep’s meat, liver and kidney contamination by heavy metals such as: copper, lead, zinc, cadmium and cobalt in different areas of al Sulaimanyah Governorate in comparison with international allowed levels. For the above purpose; three samples of (meat, liver and kidney) were taken in three different

districts of al Sulaimanyah Governorate were covered: Said Sadiq, Dokan district and sulaimanyah city center. The samples were collected during October and November 2020. The triple interference (factors) has affected significantly, the amount of copper in the different sheep tissues; so the amount was varied with the difference of tissue, the place and the time of taking the sample. The highest level of copper in Liver’s tissue was recorded in Dokan district during November, while the lowest level of copper in the meat tissue was recorded in Said Sadiq district during November. The triple interference for the study factors, also affected the level of Zinc in different sheep tissue were the amount varied by tissue difference, place and the time of sample taking. Highest level of Zinc was recorded in kidneys tissue, in Sulaimanyah city Centre during October, while less amount of Zinc was recorded in liver’s tissue in Said Sadiq during October. The triple interference within the study’s factor, significantly affected the amount of cadmium. The amounts were varied by difference of tissue, place and time of taking the samples. Highest level of cadmium was recorded in the meat tissue, at Sulaimanyah city Centre during October, while less amount of Cadmium was recorded in liver’s tissue, Said Sadiq district during October. The triple interference did not affect significantly the amount of Lead and Cobalt in different sheep’s tissue.

I'm trying to work out which cell culture medium is the most appropriate to culture these 2 cell lines. I have never worked with non-cancerous cell lines, and I am asking myself if there are easier culture protocols that the ones stated in the ATCC web page.

Has anyone used different mediums than the ones in ATCC or has a protocol they are willing to share?

Hello everyone,

I am working with Precision Cut liver slices of 250 µm. These need to be processed for FFPE. As you can imagenie they are very fragile and tend to break when I embedd them in Paraffin after fixation and dehydration.

Does someone have experience embedding thi tissues in HistoGel before Processing? My Plan is to fix the slices in Formalin over night, then embedding them in Histogel followed by processing for FFPE. Would it make sense to fix the HistoGel, too?

How does HistoGel behave when cutting?

Thanks in advance :)

Best, Franziska

Dear All,

Can microsomes be obtained from liver tissue without ultracentrifugation for in-vitro drug metabolism?

Thank you.

My lab has one or two vials that we obtained from a professor that left, but we'd like to get a more official stock. Are there any companies selling them?

I have seen some dark areas in my sirius red staining of CCl4 treated mouse liver.

However, I am not sure what are these areas indicating.

It would be nice if some histology expert could help me identifying them.

Thanks a lot in advance.

Many patients gets liver necrosis, and other liver toxicity suspected to have come from prescribed drugs mainly anti-drugs. There are many causes but specifically I wanted to localize on because it has risen many questions even from local population after being initiated to those drugs they developed liver ailments.

Mass (kg) X acceleration = Inertia which is the same as the intersection of the base. The product of inertia if we multiply it by the height we find the overturning moment of the column. If we have a wall that has a double lever arm (except for the lever arm of height and that of width) then the product of the tipping moment is divided by the width of the wall and this will be the tipping moment of the wall. If the wall is anchored at its base, a reaction will be created to the overturning torque of the lever, which multiplies (as we have seen) the overturning forces, since, as the height increases, its overturning force also multiplies. If the anchor is at the bottom of the wall, the critical failure area will also appear there and the anchor point is also the lever of the wall. Question If the anchoring of the wall is not at its lower ends, but is at its upper ends. That is, if we place this wall on a machine - press and apply pressure to it, it will remain a lever arm or its mechanical condition will change; 1) Will we have a multiplication of the tipping forces as it happens when the anchor is applied to its lower extremities? 2) Will a critical area of ​​failure of right forces N (compression and tension) be created as it happens when the anchorage is applied to its lower extremities? In short, we know that the walls drop high torques at the base since that is where the reaction of any substandard anchorage is. If the anchoring is done on the roof (ie if pre-tensioning is applied between the upper ends of the sides of the wall and the foundation ground) it will lower torques at the base and will create or not a critical failure area;

Just for some context, I'm isolating non parenchymal cells with Pronase+Collagenase D, which is a well known protocol in my lab. I use DAPI as a viability dye in my FACS panel but I encounter some issues as it also stains high DNA content cells or mitotic cells, representing around 20% of the cells (see picture). No solution in the protocol contains detergent that could permeabilize the cells, and the mouse used is a classic healthy C57 so mitotic cells are not expected, at least to that extent. Did anyone encountered the same issue in the past? Thank you

I fed this mouse normally for 6 days (control group). When I dissected this mouse, I found a white ball on the surface of its liver. What is it?

Hello!

How can the total amount of lipids in the liver be biochemically determined without the use of chloroform and methanol?

I am currently amplifying a 2kb fragment from genomic DNA from mouse liver tissue. I am not sure if the DNA sample is of good quality or maybe it is even broken into fragments.I learned that I can check this with a gel elctrophoresis. How much DNA and loading dye should I add in the well?

I would like to extract DNA from animal tissues like Liver or Spleen and I'm searching for the best kit that can give a good and pure DNA product.

Hello everyone!

I need to grow human liver cells (more specifically hepa RG cells) in a CO2-independent manner. Until now, I have used a Williams medium to grow them, but it has a bicarbonate buffering systems so it requires a controlled CO2 atmosphere. Does anyone have experience in growing these or other hepatocyte cell lines in CO2-independent media such as the Leibovitz L15 medium?

Thank you for your help!

Hello researchers,

This is my first time working on monocytes, and I plan to deliver a drug to liver monocytes. Could you please suggest to me a way deliver this drug specifically to the liver monocytes?

I have considered exosomes and hydrogels, but I don't know how they specifically deliver to the monocytes! I am looking forward to your experience on this topic.

All the best.

I am working on qPCR of cDNA from liver in various states of health and disease, and our housekeeping genes show a fair amount of variation between disease states and other groupings. Which housekeeping genes do you use for liver research?

I am currently doing a literature review, which is partly on fat-soluble vitamin metabolism in chickens. However, almost all recent research I can find on fat-soluble vitamin absorption & metabolism is focused on mammals. I was wondering how comparable fat-soluble vitamin absorption & metabolism is in e.g. humans vs. chickens. Is research done with Caco-2 cells or in mice, for example, translatable to chickens? Additionally, does this differ between parts of the body? Is there a big difference between how strongly e.g. enteric absorption and liver metabolism are conserved?

For patients taking Glutathione supplementation for Ulcerative Colitis or other disease, will this at all negatively impact the liver's ability to produce it's own Glutathione?

Hi all,

Now, I am doing quantification of infiltrated leukocytes in the liver by flow cytometry and qPCR.

Is perfusion needed before I harvest mouse liver?

Are intra-sinusoidal leukocytes "infiltrated-leukocytes"?

If so, I wonder liver perfusion washes out most of leukocytes in the liver sinusoids.

Thank you for your help.

Sincerely,

Park

Looking for a normal liver line to compare drug toxicity with HCC lines.

Would I be able to do immunofluorescence on my liver tissue if I do not perfuse before harvesting? I don't intend to use the 488 channel.

I have a batch of fish liver samples than I would like to preserve for internal controls during EROD assays. Would lyophilizing the fish liver samples prolong the length of time that I can keep them stored -80C without affecting EROD activity?

Persistence of Hepatitis B surface antigen (HBsAg) in the serum is one of the defining features of chronic hepatitis B. Regardless of the exact phase and the eligibility for treatment, many patients with chronic hepatitis B are concerned whether their blood test will clear after treatment. Therefore, can it really clear?

If so, which specific treatment is documented to do that? Even after clearance of the viral marker, does the risk of liver damage decrease?

I recently did WB to check the pCREB/CREB after glucagon stimulation in mouse liver, but I got no band both for pCREB and CREB. I used RIPA buffer with proteinase and phosphorylase inhibitor cocktail to do the liver protein extraction. Mix the liver lysates wtih Lameli buffer and BME, and run with reducing SDS-PAGE. Does any one ever have the experience to do the WB for pCREB and CREB in mouse liver? Do I need to do the nuclear extraction?

I am not sure if the problem is the specifity of the antibodies or the sample preparation.

Both pCREB (9198) and CREB (9197) antibodies are from CST.

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

Hello

I want to measure the amount of HDL in the liver, but the existing formulas show me negative values. Is this normal?

Is there a specific formula for measuring HDL in the liver?

I triglycerides and LDL cholesterol levels in've got

Hi!

I want to know what is the best method to lysis rat liver tissue in western blot method. I am using RIPA buffer and I am not getting results. Do you think we can use Rıpa buffer for liver tissue? As a result of my research, I saw that the Rıpa buffer is used, so I use the Rıpa buffer. The recipe for my ripa buffer: 150mM Nacl, 50 mM Tris (Ph 8.0) 1% NP-40, 0.5% Sodium deoxycholate, 0.1% SDS. I use 500 ul of ripa buffer and 5 ul of protease and phosphatase inhibitor cocktail per 10 mg of liver tissue. Then I sonicate at 45% amplitude 3 times at 10-second intervals, for a total of 30 seconds. Then I centrifuge at 14000 rpm +4 C for 20 min. Do you think there is something wrong with my method? Thank you from now.

Didem KORKMAZ

In our mouse bladder and liver sections we see a great deal of folding (creases in the tissue coming out of plane from the slide). May be noticed as early as upon transfer to water bath: tissues of liver section appear to wrinkle or crease almost like shrinking (43C DI water). Next day resting slide for 10 minutes at 65C does not seem to significantly decrease folding.

Sectioning method uses paraffin block chilled in freezer and regularly chilled with ice pack. Regardless of new or worn blade portion.

Surface guided radiotherapy is the state-of-the-art imaging module for patients who underwent for the radiotherapy. Some commercial available SGRT modules work on the principle of rigid registration algorithm and others work on the basis of DIR algorithm for surface matching. So, which algorithm is best to use in clinics for SBRT of liver cases?

I am using an enzymatic kit for Lipid analysis from liver tissue and I have trouble in converting the 0.018 mM of TG to mg/g of liver.

Please help me with this. The details of the experiments are given below.

Weight of liver tissue used =106 mg.

Briefly, the tissue was extracted using chloroform:methanol and filtered to make upto 6 mL volume with the same solvent.

Then, 250 μL was taken from it and added 250 μL 1X triton-100.

the solution was allowed to dry under Nitrogen gas and after complete evaporation of the solvent the residue was dissolved in 0.5 mL of distilled water (considered as the final volume for calculation by me).

50 μL of samples were used for the enzymatic assay.

The molecular weight of TG= 885.4 g/mol (as given in the kit manual)

I tried to follow the equation mass (mg)=concentration (mM)* volume (mL)* molecular weight (g/mol), which give an answer 75.5 mg/g of liver.

But I don't know if this is the right method.

Please help me with a detailed calculation if possible.

If I had made any error in this calculation please point that out.

Thank you.

I really appreciate and respect your expertise in this calculation things.

Pleas.. what is liver important on coronavirus .this question is clear....ok

I ask and I need known physiology in liver on virus and killed him

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

hi,everyone:

I try to analysis cholesterol concentration in serum and liver recently , I see protocol and it says that the serum lipid extract sovlent do not need to add triton, but liver do.

I am curiously about why there are different?

Also, l analysis triglycerides concentration in serum and liver, both do not need to add triton.

l can’t understand why triglycerides do not need to add triton?

sorry for my poor grammar, i hope you guy s can understand what i mean 😢

I have frozen sections of 4%PFA drop-fixed, 30% sucrose dehydrated fetal livers from E14.5 mouse embryos. I sectioned them both at 10 and 5 microns. When I performed the staining on the 10 micron sections the DAPI was extremely dense and therefore I went thinner on the next round. However in both attempts I could not detect any c-Kit signal despite confirming the antibody does work on whole embryo sections. I am using c-Kit eBioscience cat. 14-1171-82

My questions are:

1. Is it just a matter of hit or miss? To my understanding there is not a robust amount present in the fetal liver at any point in time so maybe it is just a matter of staining enough slides to catch some c-kit positive cells?

2. Is there something extra needed to be done as far as the tissue prep? Most protocols in the lit are very vague. The fetal liver is full of blood cells and maybe I should consider perfusing, but then is it possible I also end up flushing out my cells of interest (HSC)?

Thanks in advance!

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that were then crossed. For future planning, is it reasonable to expand a Cas9 KI+/+ Cre+/+ colony and then breed these animals to a Cas9 KI+/+ Cre-/- mice? Then all pups will be Cas9+/+ Cre-/+. The experiment require a large quantity of male mice.

We used enzymatic digestion method (collagenase 4 & trypsin ), but the cell number was less, around 2 X 10⁵. Since flow cytometric analysis that we are doing need more events, there is no other way other than increasing the number of cells. Is there any better way to do the digestion? I found that hyaluronidase can improve the cell number if used for digestion. Can anyone suggest an alternative?

There is inverse relationship between protein intake and risk of heart diseases because cholesterol start to increase as liver and cell process fats have low efficiently ( liver and other other cell required protein to its function)

Hello!

I've stained some bright field Arginase-1 Hematoxylin slides, but I'm having a hard time finding any computed histology predicates for evaluating the intensity of the cytoplasm verses the nuclei; as everything seems to be done manually.

I've attached some 20x images of the slides. But I'm concerned about edge detection of the cell and haven't had much luck with segmentation in ImageJ. I lose a lot of edges in-between some of the cells.

Thank you for your responses.

I looking for answer to above question

All comments are respect

Yours sincerely

Dear researchers,

I will look at ER stress proteins from rat liver tissue by Western blot method. I found the my protein concentration to be 86.386 µg / ul. How many ug of protein should I choose accordingly? How many ul of Laemmli Buffer Solution (2x) should I add for loading? Is the concentration of the liver lower compared to other tissues? Should I be careful about this when determining the protein amount?

Thank you.

Hello,

are there any murine cancer cell lines out there that once injected subcu will metastasize to the liver? I know there is LLC and some melanoma cell lines that will go to the lung. Appreciate any input!

thanks

I want to do tissue subcellular fractionation of mouse liver. According to the method, the speed to separate ER is over 200,000g for 120 min (Song et al., 2006). However, liver exosome isolation only needs 100,000 x g for 70 min (Ishiguro et al., 2019). Thus, I'm wondering how to separate exosome and ER?

Song, Y., Hao, Y., Sun, A., Li, T., Li, W., Guo, L., ... & Wei, H. (2006). Sample preparation project for the subcellular proteome of mouse liver. Proteomics, 6(19), 5269-5277.

Ishiguro, K., Yan, I. K., & Patel, T. (2019). Isolation of tissue extracellular vesicles from the Liver. JoVE (Journal of Visualized Experiments), (150), e58649.

Hi,

I have been doing Reverse Transcriptase qPCR on some liver mRNA samples. I am targeting liver inflammation genes like Col1a1, alpha SMA, IL6, IL1B and etc... Even though mRNA concentrations are good and primers are working, some samples are not working (difference between two samples are higher than 0.5) for a few runs in a row. After I started using triplicates of repeat samples instead of duplicates, I am getting more successful results. For analysis, I am considering the closest two and taking the average of these two samples. Does it sound appropriate in terms of data analysis ? Has anyone been doing this ?

Hi, im newbee for analysing gen expression data set and hope someone have som advise.

Im working on mice liver gen expression (mRNA) data sets generated with EdgeR - now I want to do some gen enrichment analysis - but is confused as to which methods are most robust and reliable (and to some extent user-friendly).

I have tried EnrichR, ROntoTools and GSEA - out of these, I think GSEA is the most difficult to understand.

So - what do you use? Do you have any tips for me?

I am working on liver disease classification using Machine learning. I would like to know is there any public dataset available for liver disease patients data which includes both liver functional test results and liver image of same patient.

Dear colleagues,

I would like to ask you if anyone has experience with the fixation of whole rat organs (heart, lungs, liver, kidneys, brain, testes) in 10% buffered formalin.

Thank you,

Roman

Which integrin subunit plays more important role in the liver? In connection with TGGB Community Verified icon Community Verified icon