Bio-Rad ReadyPrep™ Reduction-Alkylation Kit - Manual (Page 7)

Section 4
Instructions for Use
Per 500 µl sample volume:
Note: This protocol can be scaled for smaller and larger
protein sample volumes by adjusting the amounts of
alkylation buffer, TBP, and iodoacetamide solution
accordingly.
1. Prepare the iodoacetamide solution immediately before
use by dissolving the contents of one vial in 0.6 ml of
ultrapure water. Bio-Rad ReadyPrep proteomic grade
water (catalog #163-2091) is recommended. If not
prepared previously, open an ampoule of TBP and
transfer the solution into one of the empty glass vials
provided. Keep both solutions on ice.
Note: The iodoacetamide is unstable in solution and
therefore requires preparation immediately before use.
2. Prepare a protein sample in the desired rehydration/
sample buffer (refer to Section 5.1 for recommenda-
tions on buffer recipes and volumes). The reduction-
alkylation reaction can be performed on a
concentrated protein sample (up to 10 mg/ml) and
diluted later with additional rehydration/sample buffer
for loading onto IPG strips.
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