Does TCEP react with maleimide?

These results indicate that both TCEP and THPP react with maleimide at approximately the same rate as the addition of maleimide to the cysteinyl thiol.

Is maleimide conjugation reversible?

Since disulfides do not react with maleimides, it may be necessary to reduce these disulfide bonds prior to conjugation. Conjugation is also reversible, so it is imperative that the buffer used is made and maintained between pH 7–7.5.

Does maleimide react with cysteine?

Maleimide groups typically react with the thiol side chain of cysteine residues, which exhibit a pKa ~ 8.4. To maximize the specificity and reactivity of the cysteine–maleimide reaction, conjugations are typically carried out between pH 6.5 and 7.5 (Hermanson, 2013).

What is maleimide used for?

Maleimides linked to rubber chains are often used as flexible linking molecules to reinforce rubber in tires. The double bond readily reacts with all hydroxy, amine or thiol groups found on the matrix to form a stable carbon-oxygen, carbon-nitrogen, or carbon-sulfur bond, respectively.

How do you inactivate TCEP?

Inactivation is achieved by adding reducing agents like TCEP or DTT, where TCEP* is the recommended reducing agent. The inactivation protocol can be adapted to several workflows by varying incubation time, temperature and concentration of the reducing agent.

Is TCEP a reducing agent?

Thermo Scientific Pierce TCEP-HCl is a potent, versatile, odorless, thiol-free reducing agent with broad application to protein and other research involving reduction of disulfide bonds.

Is maleimide aromatic?

The heated benzocyclobutene is in equilibrium with the dienic system (74) which directly gives the aromatic imide (75) by Diels–Alder reaction with maleimide.

Which amino acid would Iodoacetate react with?

Iodoacetate (and bromoacetate) can react with several nucleophilic functional groups within proteins. Their relative reactivity toward protein functionalities is sulfhydryl>imidazolyl>thioether>amine.

Is maleimide soluble in water?

Both complexes containing the 6-maleimido-4-oxacaproic ester (15, 16) showed good water solubility (≥8 mg/mL) and CE experiments revealed rapid binding to human serum albumin and the formation of biadducts with dGMP and dAMP.

Is TCEP stable in water?

TCEP is stable in aqueous, acidic, and basic solutions. When TCEP is dissolved directly in water, the resulting pH is approximately 2.5. TCEP is not particularly stable in phosphate buffers, especially at neutral or alkaline pH.

Does TCEP break disulfide bonds?

TCEP is an effective reagent for the cleavage of disulfide bridges.