Reduction and Alkylation (optional)
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Dice gel slice with a fresh razor blade to bits ~2 mm x 2 mm.
-
Wash gel slice 30’ in 1 ml 30% MeOH, discard wash.
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Wash gel slice 60’ in 500 ul 100mM ammonium bicarb, discard wash.
-
Add 160 ul 100 mM ammonium bicarb that contains 2.5 mM DTT (2.5
ul 1M DTT/ml bicarb), inc 30’ at 60C.
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Add 10 ul 100 mM iodoacetamide (18.5 mg/ml in water; final [] = 5.8 mM), inc RT 30’ IN DARK, discard solution.
Reduction and
Alkylation / Digestion / Extraction
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