Bio-Rad ReadyPrep Protein Extraction Kit (Signal) 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 final pellet containing the signal proteins. 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)
To 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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