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Why is protein oxidation hard to detect?
It is unstable and artificial oxidation occurs during lysis
Why is acid lysis used?
Low pH keeps cysteines protonated and reduced
What chemical selectively binds reduced cysteine?
Cysteine-binding reagents
How does DTT help detect disulfides?
It reduces disulfide bonds
What indicates oxidation in DTT-sensitive assays?
Shift from dimer to monomer
What are limitations of disulfide detection?
Mixed disulfides and need for specific antibodies