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Questions related from Devi Prasad V
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...
02 February 2020 2,548 1 View
During transfection of B cells with plasmids/linear DNA fragments, do the large amounts of foreign (bacterial) DNA sequences activate signalling cascades, like that of TLR9 for CpG-rich sequences...
08 August 2018 2,306 1 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...
07 July 2017 7,029 0 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...
01 January 2017 7,850 11 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...
07 July 2016 6,469 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...
06 June 2016 2,066 4 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...
11 November 2015 8,292 11 View
I am interested in making a bicistronic system using viral 2A peptide to clone my protein of interest. The usual order of cloning is GOI-P2A-GFP (for example). Since the cleavage site is at the C...
08 August 2015 8,752 2 View
I have observed multiple times that if I use a PCR amplicon (with / without PCR cleanup) as a template for a subsequent PCR with the same primers, I get a smear on agarose gel. The size of the...
05 May 2015 9,554 20 View
I have been resuspending protein isolated by Trizol reagent in 1% SDS and storing at -20 degrees C. But everytime I need to use the protein for quantification, electrophoresis, etc., I need to...
05 May 2015 6,888 2 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...
02 February 2015 3,004 8 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?
09 September 2014 4,109 10 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...
09 September 2014 7,704 8 View
I am doing an assay for Carbonic anhydrase, with Sodium Bicarbonate as substrate. I find that the non-enzymatic rate itself is high, and hence am not able to detect a much steeper activity with...
03 March 2013 9,753 7 View
Can the T and B cells be inhibited for a particular antigen alone? if yes, how?
05 May 2012 4,983 8 View
Most of us use Ethidium Bromide in labs. Are there any microbes that are known to degrade EtBr? If not till now, is it possible to produce such an organism?
05 May 2012 8,680 90 View
