The new ChromaLink™ Biotin 345S is faster and easier than the traditional and destructive HABA:Avidin displacement biotin measurement method. Although widely used by researchers labeling antibodies and other molecules with biotin, the HABA:avidin method for estimating the extent of biotinylation has limitations. Because there are not many other methods available for estimating the extent of biotinylation, researchers using the HABA:avidin method have learned to live with some disadvantages. ChromaLink™ Biotin 345S accurately determines the degree of biotin incorporation without any of these disadvantages.
This protein-labeling reagent, also available in an easy-to-use kit (MVS-B-9007-105), contains three important features that make it ideal for biotinylation:
- A chromophore that absorbs strongly at 354 nm when coupled to protein; the chromophore, also covalently attached to the protein, is used to estimate the extent of labeling
- A long polyethylene glycol (PEG3) spacer, which reduces the tendency for aggregation of the labeled protein and minimizes steric hindrances related to the binding of biotin to biotin-binding proteins in affinity or detection applications
- An aromatic N-hydroxysuccinimide (NHS) ester reactive group, known to efficiently couple to amines, such as the e-amino group of lysine (K) or the a-terminus of the protein in aqueous systems
- Biotinylate protein or peptides
- Quantitate the extent of biotin labeling
- Optimize biotin-avidin complexes
- Fast and easy method to biotinylate and determine the amount of labeling
- Label and determine the extent of biotin labeling with one reagent
- Simple calculations based on UV absorbance estimates total biotin incorporation (moles of biotin per mole of protein/peptide)
- Nondestructive assay does not destroy biotinylated sample
Kit version contains sufficient material to biotinylate 5 – 50 mg of protein and delivers 85% labeled protein recovery with essentially no contamination from non-reacted or hydrolyzed reagent.