Digestion
-
Add 500 ul gel wash buffer (50% ACN/50 mm Tris pH 9.2 (for Endo-Lys C) or 50% ACN/50 mM Bicarb (for Trypsin), 60’ RT, discard wash
-
Wash gel slice with 50 ul ACN, let stand 10’, discard
-
Speed Vac to dryness without radiant cover (~15 min)
-
Add 10 ul of 100 mM ammonium bicarb containing 0.2
ugm
Promega modified Trypsin or 10 ul 100 mM Tris pH 9.2 containing 0.2
ugm of Waco EndoLysC; allow to absorb
-
Add additional 100 mM ammonium bicarb or 100 mM Tris pH 9.2 (without enzyme) to allow slices to swell to original size plus 20
ul additional
-
Incubate overnight at 40ºC
Reduction and
Alkylation / Digestion / Extraction
/ Print
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