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 scanner for signal detection) and not bad on cost. And it does detect hexa- and decahistidine tags. Not using HRP anymore since we switched over to IR detection a few years ago.
HTH- Brad On Tue, Jun 26, 2012 at 4:17 AM, Luca Jovine <[email protected]> wrote: > 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: > > > http://www.qiagen.com/products/protein/detection/qiaexpressdetectionsystem/penta-hishrpconjugatekit.aspx > > but we have never tried it... and now I'll stop here, otherwise you'll > start thinking I own QIAGEN shares :-) > > Luca > > > On Jun 26, 2012, at 09:54 , Juha Vahokoski wrote: > > 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? > > Does anyone know a good HRP conjugated antibody? > > Regards, > Juha > > On 26 June 2012 06:59, Luca Jovine <[email protected]> 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): >> >> >> http://www.qiagen.com/products/protein/detection/qiaexpressdetectionsystem/penta-hisantibody.aspx >> >> Good luck, >> >> Luca >> >> ---------------------------------------------------------------- >> Luca Jovine, Ph.D. >> Assistant Professor & EMBO Young Investigator >> Karolinska Institutet >> Department of Biosciences and Nutrition & Center for Biosciences >> Hälsovägen 7, SE-141 83 Huddinge, Sweden >> Voice: +46.(0)8.524-81136 FAX: +46.(0)8.6081-501 >> E-mail: [email protected] >> W3: http://jovinelab.org >> ---------------------------------------------------------------- >> >> ________________________________________ >> From: CCP4 bulletin board [[email protected]] on behalf of D Bonsor [ >> [email protected]] >> Sent: Monday, June 25, 2012 23:33 >> To: [email protected] >> Subject: [ccp4bb] Off-topic His-Antibody >> >> Are there any good antibodies for His-tags on the market. I have never >> used one and I heard several stories that they were poor and not worth the >> money, over the past few years. The only post I could find on the BB was >> from 2008. Have His-Antibodies improved or are they still rubbish. >> >> >> >> > > > -- > "Genius may have its limitations, but stupidity is not thus handicapped." > -Elbert Hubbard > * > > >
