In article <60j6uk$pbp at dfw-ixnews10.ix.netcom.com>,
babco at ix.netcom.com(Thomas R. Anderson) wrote:
> In <342AB14C.CD4997A0 at med.unc.edu> syan <syan at med.unc.edu> writes:
> >
> >Has anyone out there had any success using anti-His tag antibodies for
> >western blots? <snip>
>> With obvious bias, I might suggest that you ours. We have only had the
> antibody on the market for a couple of months, but have heard from a
> number of our customers that they are getting cleaner results with this
> antibody compared to whatever they were using before.
>> Tom Anderson
> Berkeley Antibody Company (BAbCO)
> tel: 510-412-8930 or 800-922-2226
> fax: 510-412-8940
> email: tanders at babco.com> www.babco.com
Blatant advertizing !
Well, then I guess I might share my experience of the BAbCO anti His
antibody with the group, as a bit of unbiased feedback. First of all we
had to buy quite a large aliquot (for ca 500$) for testing it out. Well,
fine, here are the results. By the way, everything detected using the
Pierce version of ECL.
The antibody does indeed recognize most of the His-tagged proteins that I
tried it with. Unfortunately it did not recognize the C-His-tagged
protein that I most urgently needed to detect. I loaded 200 ng of Ni-NTA
purified protein on a western and there was no signal.
It can be used to detect highly expressed proteins (from baculo or E.coli)
but with HeLa cell extracts I had high background. The Carbonic anhydrase
band in my markers also shows a very strong signal!
Finally we got some Qiagen 5His ab from a neighboring lab and compared
side by side with the BABCO antibody. The results were quite clear. The
Qiagen stuff gave much stronger signals and lower background. Tested with
three different his-tags (6His-N, 10His-N and 6His-C) on PCNA.
The documentation from Qiagen is also much better and they claim that
their Ab can be used for IP's as well. I didn't compare the price
however.
Conclusion:
This antibody works for most proteins but has some background problems
and is probably not the best anti His antibody on the market.
If sincerely hope that others have had better luck with the BAbCO antibody!
Zophonias
PS. I have no connections with either BABCO or Qiagen except as a customer.
_____________________________________________________________________
Zophonias O. Jonsson
Institut fur Veterinarbiochemie Tel: (41-1)-635-54-75
Universitat Zurich-Irchel Fax: (41-1)-635-68-16
Winterthurerstrasse 190
CH-8057 Zurich
Switzerland
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