Does maleimide react with lysine?
From pH 6.5 to pH 7.5, the thiol-maleimide reaction is chemoselective for thiols. (a) The reaction is chemoselective for thiols from pH 6.5 – 7.5. (b) Above pH 7.5, thiol chemoselectivity is lost, and the maleimide moiety begins reacting with free amines (e.g., lysine).
Does maleimide react with amine?
Maleimide reaction chemistry In more alkaline conditions (pH >8.5), the reaction favors primary amines and also increases the rate of hydrolysis of the maleimide group to a non-reactive maleamic acid.
Is maleimide conjugation reversible?
In Brief Download the PDF of this protocol 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 TCEP react with maleimide?
TCEP can keep the cysteines from forming di-sulfide bonds and unlike dithiothreitol and β-mercaptoethanol, it will not react as readily with the maleimide. However, TCEP has been reported to react with maleimide under certain conditions. TCEP is also used in the tissue homogenization process for RNA isolation.
What is maleimide soluble in?
|Molar mass||97.07 g/mol|
|Melting point||91 to 93 °C (196 to 199 °F; 364 to 366 K)|
|Solubility in water||organic solvents|
How do you dissolve maleimide?
Dissolve maleimide in DMSO or fresh DMF (1-10 mg in 100 uL). Add dye solution to thiol solution (20x fold excess of dye), flush vial with inert gas, and close tightly. Mix thoroughly, and keep overnight at room temperature, or 4 Celsius.
Is maleimide toxic?
Toxic if swallowed. Causes severe burns. Risk of serious damage to eyes. May cause SENSITISATION by skin contact.
Is TCEP nucleophilic?
TCEP is stable in aqueous solutions, highly reactive, and selective towards disulfide structure. Nucleophilic Substitution by the Phosphorus Atom of TCEP.
Does 280 absorb TCEP?
Moreover, TCEP does not absorb UV light in the range of 250 – 280 nm, making it useful for monitoring reactions with proteins that contain aromatic amino acid residues.