Bio-Rad ReadyPrep™ Protein Extraction Kit (Membrane I) User Manual
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5.2 Preparation of 2-D Rehydration/Sample Buffers
The PSB and PSB diluent are provided with this kit to
solubilize the protein pellet after using the ReadyPrep 2-D
cleanup kit. PSB is a proprietary, strongly chaotropic 2-D
rehydration/sample buffer that will solubililze both
hydrophilic as well as hydrophobic proteins. Other strongly
chaotropic 2-D rehydration/sample buffers (Section 5.2.2)
may be substituted for PSB or used for diluting an
appropriate amount of sample for IEF/2-D analysis.
5.2.1. Complete Protein Solubilization Buffer (PSB)
o make 2 ml of complete 2-D rehydration/sample buffer,
add 1.1 ml of PSB diluent to each 1 g of PSB powder.
Note: Before weighing out the PSB powder, shake the bottle
vigorously 10-15 seconds to break up any clumps and to
ensure a uniform blend of the different components.
Mix the solution until the powder is completely dissolved
(the tube can be warmed to speed dissolution of the solids,
but do not allow the temperature to exceed 30°C). Add
DTT, Bio-Lyte ampholyte, and Bromophenol Blue according
to the table below to complete the preparation of the
buffer.
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