CoA-SH is a maleimide-reactive building block which can be used for the one-step synthesis of CoA substrates from maleimide-containing precursors. This building block allows you to make custom CoA substrates for labeling ACP-tag or MCP-tag fusion proteins for a wide range of applications.
A typical reaction is performed at 30°C overnight by using N,N-dimethylformamide (DMF) and Tris-buffer (pH 7.5) as solvents. Equimolar quantities of CoA-SH and reactive maleimide are recommended.
Example reaction: Coupling of CoA-SH to a Maleimide-reactive Precursor
Figure 2. Coupling of CoA-SH to a maleimide-reactive precursor. A solution of a maleimide derivative (1 eq, 6.6 μmol) in DMF (450 μl) was added to a solution of CoA-SH (6.2 mg, 1.2 eq, 7.9 μmol) in Tris-buffer (pH 7.5, 50 μl). The reaction mixture was shaken overnight at 30°C. The solvent was evaporated under vacuum. The solid was dissolved in 1 ml water/acetonitrile (9:1, 0.08% TFA) and purified by HPLC. HPLC fractions containing the product were neutralized with NH4OH (30% aquaeous solution) and lyophilized.
Save your favorite products by clicking Add to My NEB, making re-ordering and remembering what you need quick and simple.
Returning to use the same tools or tables often?
Click Add to My NEB to save the links and view as often as you like.
Find great application content?
Click Add to My NEB to save and impress your labmates with your knowledge.
Keep track of any other pages you find useful by clicking Add to My NEB.
The NEB PCR Cloning Kit allows direct cloning from any amplification reaction, with no purification needed!
The Gibson Assembly™ Cloning Kit is a robust method for assembling 1-2 DNA fragments, with a cloning vector, and ensuring high transformation efficiencies - all in a little less than 2 hours.