Pongpawan Sethanant

9 Questions 18 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 9,057 6 View

Can DTNB displace zinc bound to cysteine residues?

Dear all, My aim is to determine the number of zinc atoms bound to a protein. I came across a technique called PAR/PMPS assay which is quite accurate in determining the zinc-protein stoichiometry...

07 November 2017 2,506 1 View

What techniques could be used for quantifying the amount of zinc atoms in a protein?

What techniques could be used for quantifying the amount of zinc atoms in a protein? Some techniques that I could think of are PAR-PMPS assay and atomic absorption. I would really appreciate any...

23 July 2017 5,993 2 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 3,757 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 6,661 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 6,191 5 View

In CRISPR-Cas9 genome editing, not all genes have PAM sequence, so how do they overcome this?

As the topic implies, when we perform CRISPR-Cas9 genome editing, PAM sequence located after the gene/DNA sequence of interest is essential. However, not all DNA sequences/genes have PAM sequence...

10 February 2016 3,372 4 View

What is like to be a structural biologist in a pharmaceutical company?

I saw some structural biology positions in many pharmaceutical companies, but what do they actually do? Is it the same as doing research? Just curious. Thank you in advance for all the answers. :)

08 February 2016 3,515 1 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 2,576 6 View