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

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View