Alex Khan

4 Questions 4 Answers 0 Followers

Questions related from Alex Khan

Can I add more RIPA buffer to cell lysate if I didn't add enough during the initial extraction?

I tried running cell lysate from epidermal keratinocytes on a gel during sds-page but realized that it is severely contaminated with genomic DNA - I am fairly certain this is because I did not...

22 October 2021 9,130 1 View

Should methanol and/or SDS be included in the transfer buffer for a native western blot?

As the title suggests, should SDS and/or methanol be included in the transfer buffer for transferring large molecular weight proteins (>300 kDa)? If so, how much of each is optimal? For...

12 October 2021 2,059 1 View

Why does my western blot membrane have black spots and smears on it?

I have continually had issues with bad looking western blot images, specifically black dots and smears on the nitrocellulose membrane. Does anyone know what could be the cause of this? I have read...

01 October 2021 6,712 5 View

Western blot tips and tricks for detecting a 900 kDa protein under native conditions?

I am currently trying to detect the collagen 7 (C7) trimer protein (~900 kDa) in its native form via western blot - it is made up of three C7 monomers of roughly 300 kDa. I have already detected...

20 September 2021 7,450 10 View