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Questions related from Devi Prasad V
I have an issue in analysing qRT-PCR datasets. For my gene of interest, treatment A's mRNA fold change values are 10, 40, and 200, over the control group. For treatment B, the corresponding values...
12 March 2021 6,153 4 View
I plan to perform ChIP experiments with wild-type mouse cells (no GFP transgene) using an anti-protein antibody raised in goat. May I use an anti-GFP antibody of the same isotype raised in the...
11 February 2020 1,626 1 View
I have observed that when I pulse cells with EdU for analysing cell cycle kinetics, the pattern of staining is as in Fig.1. Considering the heterogeneity of S phase cells during the pulse, cells...
05 June 2018 9,425 28 View
I use Lectin PNA from Arachis hypogaea (peanut), Alexa Fluor® 647 Conjugate (Cat No. L32460), 1mg/ml, to stain germinal center B cells in murine splenocytes. With dilutions of 1:100, 1:500 and...
02 May 2018 7,487 7 View
I am observing cell proliferation kinetics of B cells using CFSE staining. If I stain quiescent cells with CFSE and chase them for 3-6 days, they show distinct peaks for different generations....
04 August 2017 4,867 4 View
I used 3 RIPA lysates from murine cells - from a lymphoma cell line, primary naive B cells and primary stimulated B cells. I used anti-Lamin B1 antibody ab133741 at 1:5000 dilution. I see a clear...
28 July 2017 2,018 0 View
I treat quiescent murine B cells with some stimulants to make them divide. They enter into the cell cycle and they divide for about 10 days and then stop dividing. At any point, if the stimulants...
09 June 2017 718 8 View
Against an epitope, high-affinity BCR means strong signaling and vice-versa. But what does affinity mean in terms of molecular events? How does the cell know the BCR is of high-affinity? Is it...
04 June 2017 566 4 View
I am doing a time course experiment to study cell divisions. I have carried out the experiments in dark, given the light sensitivity of CFSE. I have analysed stained and unstained samples at every...
16 January 2017 9,290 8 View
In immunology experiments, people use nitrophenyl (hapten) conjugated to a carrier protein and report antibody affinities based on their ability to bind to NPx, where 'x' is an integer.Could...
28 July 2016 582 1 View
Is it necessary to isolate nuclei from cells and use this RNA for qPCR? Or can specific primers be designed in intronic regions so that only unprocessed RNA will be picked, and use whole cell RNA...
08 June 2016 2,270 3 View
I am using a plasmid vector of about 8kb for cloning. After cutting with EcoRI and BamHI, I performed a ligation with T4 DNA ligase and transformed in DH5-alpha, and found that there were as many...
14 November 2015 4,631 10 View
I have been trying transfections with this cell line, with hardly any success. Bicistronic IRES vectors don't seem to work Electroporation has been giving low efficiencies High concentration of...
06 February 2015 6,327 7 View
Does anyone use a plasmid vector suitable for mammalian expression by non-viral transfections, and having the following features: -GFP following an IRES site and not as a fusion with the gene of...
12 September 2014 4,797 7 View
I use fetal calf serum in the media used to culture B cells. Would any of the serum components be antigenic and affect my experiments?
04 September 2014 4,014 9 View
It is said to be triggered by fungus. So, can it spread as spores to other people?
11 September 2012 4,389 6 View
Can the T and B cells be inhibited for a particular antigen alone? if yes, how?
05 May 2012 7,967 7 View
