Jason White

6 Questions 29 Answers 0 Followers

Questions related from Jason White

Does anyone have experience with IHC staining of his-tagged protein in mouse tumor tissue?

Does anyone have an IHC protocol for detection of a his-tagged protein in mouse tumor tissue (cryo, not paraffin)? I'm trying to stain for a protein that contains a 6His tag and accumulates in...

22 August 2014 6,203 4 View

Can anyone help with measuring protein concentration after PEG conjugation?

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...

06 May 2014 9,867 6 View

Addition of copper sulfate to cell culture media

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...

23 April 2013 9,515 7 View

Elution from 6His column

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 4,015 24 View

Does anyone have experience with expressing antibody fragments using the pEE12 system in NSO cells?

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...

23 July 2012 3,837 1 View

Protein purification from culture supernatant

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)....

15 June 2012 6,966 43 View