What Concentration Should TCEP and IAA Be Used for Protein Reduction and Alkylation
TCEP
TCEP is typically employed to reduce disulfide bonds in proteins, with concentrations commonly ranging from 5 mM to 10 mM. It is crucial to adjust the concentration according to the experimental requirements and specific properties of the protein under study.
IAA
Iodoacetamide (IAA) serves to alkylate free thiols post-reduction, thereby preventing the reformation of disulfide bonds. Standard concentrations range from 15 mM to 50 mM, though adjustments may be necessary based on experimental conditions and protein characteristics.
In experimental applications, it is essential to use TCEP and IAA at concentrations appropriate for the specific requirements of your study, while avoiding exposure to light and oxidation during handling.
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