Thank you all for your suggestions! On Mon, Feb 6, 2017 at 5:53 AM, Artem Evdokimov <[email protected]> wrote:
> Hi, > > In addition to HABA dye assay (which will work great but will also be > fooled by any biotin that is not conjugated) you can do: > > * quantitative MS > * TLC > * HPLC > * elemental analysis > * https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3614710/ biotin catalysis > of the N3- + I3- reaction (also fooled by free biotin of course) > * UV (but beware, biotin only absorbs strongly below 240nm so you're not > super well off there > > Artem > www.harkerbio.com > "all of our Biotin comes only from free-range gummy vitamin bears..." > > - Cosmic Cats approve of this message > > On Mon, Feb 6, 2017 at 2:03 AM, Debasish Kumar Ghosh <[email protected]> > wrote: > >> Hi Alex, >> >> In addition to Mirella's suggestion I would like to make an addition >> which might be specifically useful for you. Since your peptide has biotin >> tag, You may use HABA dye assay for the exact quatifiation of biotin (and >> thus biotinylated peptide). As far I recall, Thermo scientific provide a >> kit for this assay. The assay is simple and gives accurate results. >> >> Best !!! >> >> >> >> Debasish >> >> CSIR- Senior Research Fellow (PhD Scholar) >> C/o: Dr. Akash Ranjan >> Computational and Functional Genomics Group >> Centre for DNA Fingerprinting and Diagnostics >> Hyderabad, INDIA >> >> Email(s): [email protected], [email protected] >> Telephone: 0091-9088334375 (M), 0091-40-24749396 (Lab) >> Lab URL: http://www.cdfd.org.in/labpages/computational_functional_ >> genomics.html >> >> >> >> ----- Original Message ----- >> From: Alex Lee <[email protected]> >> To: [email protected] >> Sent: Mon, 06 Feb 2017 03:02:07 +0530 (IST) >> Subject: [ccp4bb] How to determine the concentration of biotinylated >> peptide? >> >> Dear All, >> >> Sorry for the off-topic question, I'd like to do Biacore SPR assay with >> N-terminal biotinylated peptide as ligand (to Biacore SA chip) and my >> protein as analyte. I have a question of how to determine the >> concentration >> of biotinylated peptide （synthetic peptide), if the peptide has no Tyr and >> no Trp residue, I guess amino acid analysis may not work because the >> N-terminal of the peptide is biotinylated. >> >> I'd appreciate if anyone share his/her experience on this. >> > >
