Has anyone successfully stained (intracellular staining) for T-bet with the Cytofix/Cytoperm Kit from BD Biosciences??

I isolate PBMCs from human blood and try to perform intracellular staining for T-bet (i'm interested in CD19+ cells, but for now i use CD3+ as a control). I do know there are kits that are...

22 September 2019 2,889 3 View

Anyone with DNA native PAGE experience (66bp PCR product)?

Hey guys, I have a PCR product that is 66bp long and want to visualise in native PAGE but the bands are not as clear as I imagined they would be. I am preparing 10% gels that I pre-run for 30mins...

24 April 2018 9,088 5 View

Has anyone prepared linear acrylamide before?

Hey, I was wondering if someone has prepared linear polyacrylamide (LPA) themselves before? I wanted to use it in DNA precipitation but it was far too expensive to buy whereas we already have...

05 December 2017 9,277 3 View

How to get rid of a negative control's Ct value in qPCR?

I run a negative control in qPCR, but I get a Ct value close to 25. I have already changed primers, ddH2O and even SYBR, but the problem still remains..

13 September 2017 8,236 6 View

Which is the command in order to model residual welding stresses in LS-DYNA?

I have modelled a heavily welded structure in LS-DYNA and I am using implicit method to solve it as the problem is static. Which is the correct command (keyword) in LS-DYNA in order to model...

24 November 2016 367 5 View

Chiral auxilliary cleavage?

Hi, I have trouble cleaving off the phenyl glycinol chiral auxilliary from my amino acid analogue. I have only tried oxidative conditions, (PbOAc4) varying the reaction time, temperature and...

19 September 2016 7,487 3 View

Have you any suggestions of I.C.T. usage, tools, and experiments, to monitor, access and intervene in order to raise meta-cognition abilities?

03 August 2015 1,764 4 View

Cell signaling, Signal transduction and GPCRs

I start this discussion so we can all share our favourites books on cell signaling or signal transduction. I am highly interested in GPCRs. So I will start first. "G Protein-Coupled Receptors:...

01 January 1970 9,182 4 View

Which variables affect my observation? (Statistical analysis)

I have acquired 21 observations from 21 samples (density difference percentages). For this 21 samples I have also 9 other parameters for each. I would like to do a statistical analysis to find...

01 January 1970 6,758 8 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

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?

03 March 2021 1,763 2 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

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

03 March 2021 8,890 1 View

IHC on mouse liver hepatocytes?

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...

01 March 2021 6,953 1 View

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

What are the pros and cons for mice versus rats?

Hi Everyone, I am designing a pilot study using rodents to assess the effects of simulated space environments (radiation + altered gravity) on microbiome and immune system. I have limited...

24 February 2021 1,825 3 View

Is it harmful to breed for Cre Homozygosity initially?

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...

24 February 2021 5,759 3 View