Dear Neil:
For biotin labeling please refer the published study of the Peptide
Synthesis Research Group by Bibbs et al in J Biomolecular Techniques
Vol 11, Issue 4 Dec. 2000.
For Fluorescein coupling I use the following protocol for on-resin labeling:
FITC (10 fold molar excess over peptide) dissolved in:
1ml 0.05M borate buffer pH 9.0
1ml ethanol
8ml DMF
I perform the step manually and check completeness of coupling by Ninhydrin.
If you label at a Lys side chain , incorporate the N-terminal AA with
Boc protection. That way, you don't risk Fmoc deprotection at pH 9
and you still remove the Boc group upon cleavage.
For cleavage I have used reagent B successfully.
Henriette
>Dear All
> I have got to make some peptides labelled with biotin and
>fluorescein. This will be the first time I have attmpted such labelling and
>I wonder if anybody out there in ABRF-land has any Protocols/references for
>on resin labelling of peptides with such labels as biotin/fluorescein/DIG
>etc.
>
> Many thanks in advance, Neil Brew
Henriette A. Remmer, Ph.D
Director of Protein Services
Protein Sciences Facility
University of Illinois
311/315 Noyes Laboratory, Box 62-1
505 S. Mathews Ave.
Urbana,IL 61801
Tel: (217) 333-4695 (Lab) (217) 333-3841 (office)
Fax: (217) 244-1142
homepage: http://www.life.uiuc.edu/biotech/protein_sc.html
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