Pongpawan Sethanant

5 Questions 15 Answers 0 Followers

Questions related from Pongpawan Sethanant

Does addition of IPTG before and after lowering induction temperature affect protein expression levels?

Dear all, I would like to know whether addition of IPTG before (37 degC) and after lowering induction temperature (16 degC) affect protein expression levels? *add IPTG before = add IPTG when 37...

18 October 2018 8,987 6 View

How to purify oxidation prone proteins?

Dear all, I am currently purifying a small protein which contains 5 cysteines. The purification flowis: Ni-NTA -> His tag removal by TEV cleavage -> remove TEV protease by Ni-NTA column...

02 March 2017 3,696 3 View

Why is my expressed protein differs in size in different host cells?

Dear all, I am expressing a virus protein about 9 kDa on a 16% tricine gel (150V constant). I induced protein expression with 0.5 mM IPTG and collected the cells pre- and post- induction. The...

12 May 2016 6,598 6 View

How do you separate Ni-NTA beads from cell lysate?

Dear all, I accidentally mixed Ni-NTA beads with E. coli cell lysate but has a problem with separating them. Is there any way to separate them so that I can regenerate Ni-NTA beads for...

27 April 2016 6,139 5 View

I am having problems with biotinylation consistency, does anyone have any suggestions/advice?

I am biotinylating my monoclonal antibodies at the moment. However, I have biotinylated two batches of them and the reactivity & background(non-specific binding) seems to be...

22 February 2015 2,527 6 View