Bio-Rad ReadyPrep™ Reduction-Alkylation Kit - Manual (Page 12)

5.2.1. General-Purpose 2-D Rehydration/Sample
Buffer (no reducing agent)
(8 M urea, 2% CHAPS, 0.2% Bio-Lyte ® 3/10 ampholyte,
0.002% bromophenol blue)
Component Final Concentration Amount to Make 2 ml
Urea (FW 60.06) 8 M 0.96 g
CHAPS 2% (w/v) 0.04 g
100X Bio-Lyte 0.2% (w/v) 20 µl
3/10 ampholyte**
Bromophenol Blue
0.002% (w/v) 4 µl of a 1%
(w/v) solution
Proteomic grade 1.25 ml
water
** Use an ampholyte buffer that corresponds to the pH range of the IEF
separation to be performed. For example, ReadyStrip ™ micro-range buffers
with ReadyStrip micro-range IPG strips and ReadyStrip 7-10 buffer with
ReadyStrip pH 7-10 IPG strips. Bio-Lyte 3/10 ampholyte can be used with
all other ReadyStrip IPG strip pH ranges.
5.2.2. Strongly Chaotropic 2-D Rehydration/
Sample Buffers for Highly Hydrophobic Proteins
5.2.2.1. Standard Strongly Chaotropic Buffer
(no reducing agent)
(7 M urea, 2 M thiourea, 4% CHAPS, 0.2% Bio-Lyte 3/10
ampholyte, 0.002% bromophenol blue)
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