Bio-Rad ReadyPrep™ Protein Extraction Kit (Membrane II) - Manual (Page 4)

can be applied to a wide variety of biological samples, from
animal cells and tissues to yeast, bacteria and plant
tissues. After the cells or tissue are disrupted using an
ultrasonic probe, the extract is incubated in a Membrane
Protein Concentrating Reagent. This high pH buffer strips
away proteins weakly-associated with membranes while
leaving membrane fragments with their associated integral
proteins intact. Ultracentrifugation is used to concentrate
the membranes into a pellet, and the proteins are
solubilized from the washed pellet using an isoelectric
focusing-compatible 2-D rehydration/sample buffer. The
2-D Rehydration/Sample Buffer 1 utilized in the kit is a
strongly chaotropic buffer containing the zwitterionic
detergent ASB-14, making this solution one of the most
powerful solubilizing reagents available for 2-D gel
electrophoresis. The kit also contains the reducing agent
tributylphosphine (TBP), which has been shown to be more
efficient than dithiothreitol in keeping proteins reduced in
2-D PAGE applications.
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