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Questions related from Jason White
I've conjugated a PEG4-maleimide (MW 613.66) onto an antibody fragment (52 kDa), but I'm trying to figure out how this affects A260/280 measurements on a Nanodrop afterwards. After conjugation and...
05 May 2014 7,759 7 View
I have purified a protein from a Ni-NTA column and noticed that during dialysis (same buffer used for elution, without imidazole) the protein precipitates. I've spun down the precipitant, and I...
09 September 2013 3,780 18 View
I have a protein that I've purified from a His column. I've noticed that when I do a buffer exchange of the eluted fractions into PBS (pH 7.3) and then concentrate the fractions, the solution gets...
07 July 2013 8,915 25 View
I'm trying to express an antibody fragment that is covalently held together by cysteine residues that I have added to the sequence. After running the western (non-reduced) to assess expression, I...
04 April 2013 569 7 View
I am having a gene synthesized (~1300bp) that was originally expressed in E.coli but I want to use it in a mammalian expression vector. I designed the gene and had it optimized by GeneArt from...
02 February 2013 3,003 1 View
I'm trying to create a stable cell line of NSO cells that express my protein into the supernatant. I've transfected the cells, seeded them at various densities into 96 well plates and screened the...
12 December 2012 6,298 6 View
Do these cells lines need to be grown in special shake flasks, or can they can be cultured in regular T-flasks without shaking? We don't have an incubator with a shake platform so I'm curious if I...
09 September 2012 5,182 0 View
I have a 6his-tagged protein that I am secreting into the media and eluting from the Ni-NTA column, but yields seem to be really low (recovered ~300ug/L of media). I've tried running a gradient...
08 August 2012 8,349 25 View
I've been using a lentiviral-based system with a pLenti expression vector in 293T cells and can purify the fragment, but yields are excruciatingly low (~300ug/L). There is quite a bit in the...
07 July 2012 3,233 1 View
Hello protein gurus, I'm expressing a protein (~50 kDa) that contains a 6x-His tag from 293T cells into the media supernatant The media is complete DMEM (10% FBS, 1% PSG, and 0.1% Blasticidin)....
06 June 2012 3,390 44 View
