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Procedure of diagonal electrophoresis
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Pamela Anne Trinidad
Procedure of diagonal electrophoresis
You probably don’t need to oxidize the disulfide bonds rather than reduce them. Performic acid has some potential advantages:
The oxidation products can’t spontaneously reform a disulfide bond whereas the reduction products from a thiolate reductant can.
Oxidation by performic acid causes a significant mass change for the cysteine residues which can be useful for downstream mass spec. This article states that complete oxidation of the cystine to cysteic acid occurs (-SO3-), which is a molecular mass change of 48 u. With BME or DTT reduction, you are left with a thiol or thiolate.
These oxidation products are also negatively charged which can increase mobility in PAGE and possibly aid separation.
You probably don’t need to oxidize the disulfide bonds rather than reduce them. Performic acid has some potential advantages:
The oxidation products can’t spontaneously reform a disulfide bond whereas the reduction products from a thiolate reductant can.
Oxidation by performic acid causes a significant mass change for the cysteine residues which can be useful for downstream mass spec. This article states that complete oxidation of the cystine to cysteic acid occurs (-SO3-), which is a molecular mass change of 48 u. With BME or DTT reduction, you are left with a thiol or thiolate.
These oxidation products are also negatively charged which can increase mobility in PAGE and possibly aid separation.
You probably don’t need to oxidize the disulfide bonds rather than reduce them. Performic acid has some potential advantages:
You probably don’t need to oxidize the disulfide bonds rather than reduce them. Performic acid has some potential advantages:
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