good hexa-his antibody supplier wanted

Hi all,

I am working with 6XHis-tagged proteins for Ninta pulldown assays. I
subcloned a library of genes in this His vector that I generated by
oligocloning of Met-6Xhis encoding oligos. So, the His is at the
N-terminus and just 6XHis. Unfortunately, I have severe troubles to detect
my proteins with either a penta-his AB (qiagen) or a AB against 6XHis (Santa
Cruz, 16-002). I know that the cloning and expression is OK as the pulldown
works fine and I can detect the protein by the AB against the protein. but
for most proteins in my library i do not have a specific AB.

It seems that most people generated a larger tag that contains some
additional amino acids to generate a stronger epitope. We have a good his ab
that recognizes c-terminal his-fusions (it requires a his-cooh) but this is
not functional for this library of course.

Is there anyone out there that knows of a good working 6XHis AB which also
recognizes plain 6XHis fusions at the N-terminus of the protein of interest.

Thanks!!!!


Jurre

Jurre Hageman wrote:


I have used anti-His from Qiagen. They have 3 anti His antibodies,
maybe one would work for you:
[Only registered users see links. ]

NAYY
Allison

allisonh <[Only registered users see links. ]> wrote:

I'e used Qiagen's anti-5His and on Western it performed with less
background than anti-6His from Sigma.

DK