- Cell Biology
- Product Type
- Research Area
- New Products
- About Tocris
- Contact Us
This is intended as a guide only; for full experimental details please read the reference provided.
Amine reactive CoraFluor reagents contain pentafluorophenyl esters (pfp esters) which can be conjugated to (non-protonated) aliphatic amine groups. The primary reactive species for protein amine-conjugation are the ε-amino groups of lysine residues. To avoid protonating these groups, it is important to perform the reaction at a slightly basic pH. In addition, buffers containing primary amines should be avoided, since they will compete for conjugation with the pfp ester.
Please note that pfp esters can be moisture sensitive, so handle accordingly. Where possible, handle and store CoraFluor reagents in the dark.
The molar equivalents of CoraFluor can be adjusted accordingly depending on size of the protein and desired degree of labeling.
A280,corr = A280 - (A340 x c.f.)
c.f. is the correction factor for the terbium complex contribution to A280 and is equal to 0.157.
|cab =||A280,corr||x b|
where εab is the antibody/protein/nanobody extinction coefficient at A280 and b is the path length in centimeters.
|cTb =||A340||x b|
where εTb is the complex extinction coefficient at A340, equal to 22,000 M−1 cm−1, and b is the path length in centimeters.
|DOL =||cTb||x b|
TWEEN® is a registered trademark of Croda International PLC
Zeba™ is a registered trademark of Thermo Fisher Scientific
Payne et al (2021) Bright and stable luminescent probes for target engagement profiling in live cells. Nat.Chem.Biol.17 1168 PMID:34675420.