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 <dkgh...@cdfd.org.in> 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): dkgh...@cdfd.org.in, dgho...@gmail.com > 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 <alexlee198...@gmail.com> > To: CCP4BB@JISCMAIL.AC.UK > 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. >
