Bio-Rad ReadyPrep™ Reduction-Alkylation Kit - Manual (Page 11)

5.2 Preparation of 2-D Rehydration/Sample
Buffers
The 2-D rehydration/sample buffer is not a component of
this kit, but is necessary for performing 2-D electrophoresis.
It is not provided in the kit because different protein
samples can require different rehydration/sample buffers.
For convenience, a selection of 2-D rehydration/sample
buffer formulas are provided below.
Do not add reducing agent (i.e. TBP, DTT or TCEP)
to the rehydration/sample buffer before perform-
ing the reduction-alkylation reaction. A molar
excess of alkylating agent to reducing agent is
required to ensure efficient alkylation.
Users of the ReadyPrep protein extraction kit (membrane I),
catalog #163-2088, or the ReadyPrep protein extraction kit
(cytoplasmic/nuclear), catalog #163-2089, or the
ReadyPrep protein extraction kit (signal), catalog
#163-2087, should use the protein solubilization buffer
(PSB) and PSB diluent provided with these kits to solubilize
the protein samples. PSB is a proprietary strongly
chaotropic 2-D rehydration/sample buffer (refer to Section
5.2.2.2 for preparation instructions).
8
4110063A.qxd 1/29/2003 2:48 PM Page 11