Bio-Rad ReadyPrep™ Reduction-Alkylation Kit - Manual (Page 13)

Component Final Concentration Amount to Make 2 ml
Urea (FW 60.06) 7 M 0.84 g
Thiourea 2 M 0.304 g
CHAPS* 4% (w/v) 0.08 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 1.1 ml
water
*Other neutral or zwitterionic detergents can also be used at concentrations
of 1% to 2% (w/v) to improve solubilization of membrane and hydrophobic
proteins. Examples are n-octyl-
β-D-glucopyranoside, SB3-10 (N-decyl-N,N-
dimethyl-3-ammonio-1-propanesulfonate) and ASB14 (tetradecanoylamido-
propyl-dimethylammonio-propane-sulfonate).
**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.
5.2.2.2. Protein Solubilization Buffer (PSB)
PSB and PSB diluent are provided with the ReadyPrep
protein extraction kit (membrane I), the ReadyPrep protein
extraction kit (cytoplasmic/nuclear), and the ReadyPrep
protein extraction kit (signal) to solubilize the protein
sample. PSB is a proprietary, strongly chaotropic 2-D
rehydration/sample buffer that will solubilize both
hydrophilic and hydrophobic proteins. To make 2 ml of
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