Dear All,
Thanks for all the replies on and off-board. I received around twenty replies
and the majority have spoken in favor of the QIAgen BSA-free anti-5His mAb from
QIAGEN. Not to be bias, a couple of people recommended the one from Abcam as
well.
Thanks again, Dan
I have been using an HRP conjugated antibody from Thermo that can detect
maybe 3-8ng of HisTagged protein from insect cell lysates. Performance is
ok, but with the sensitivity I am not so happy with.
How is the sensitivity with the Qiagen antibody? How many picograms you
need for a clear band?
Our experience essentially agrees with QIAGEN's claim that you can detect
proteins in the low nanogram range, i.e. 1-2 ng... so not a major improvement
over your current antibody, I'm afraid. They also sell an HRP conjugate version:
I'll just second, third, and fourth the assessment of the Qiagen
anti-pentaHis antibody. I pretty much gave up on anti-His antibodies until
we tried this out in the lab and now everyone loves it. Reasonable
sensitivity and specificity (we use IR dye-conjugated secondaries and a
Li-Cor Odyssey
The one we have is rubbish. I can tell a vendor name if you want off-list.
A lot of non-specific bands + high background level makes it almost
impossible to use against cell lysate. It sort of works against purified
His-tag protein, but not perfect either.
On Mon, Jun 25, 2012 at 4:33 PM, D
We routinely use BSA-free anti-5His mAb from QIAGEN, and it works just fine (at
least using media and lysates of both insect and mammalian cells):
http://www.qiagen.com/products/protein/detection/qiaexpressdetectionsystem/penta-hisantibody.aspx
Good luck,
Luca
We also use that same antibody, and are satisfied overall.
On 6/25/12 8:59 PM, Luca Jovine wrote:
We routinely use BSA-free anti-5His mAb from QIAGEN, and it works just fine (at
least using media and lysates of both insect and mammalian cells):
