Bio-Rad ReadyPrep Protein Extraction Kit (Signal) - Manual (Page 11)

Section 5
Appendix
5.1 Rehydration/Sample Buffer Volume
In the final step for this kit, all samples are resuspended in
a 2-D rehydration/sample buffer (Section 5.2). To best
determine the volume of 2-D rehydration/sample buffer to
use, consider the questions listed below. To assist with
these calculations, the table that follows indicates
appropriate volumes of 2-D rehydration/sample buffer
needed to rehydrate IPG strips of specific lengths and the
approximate amounts of protein required for detection
using silver stain or Coomassie Blue G-250 stain. An
example illustrates how to calculate the volume of 2-D
rehydration/sample buffer required.
1. What is the quantity of protein precipitated in the tube?
2. For 2-D electrophoresis experiments using IPG strips,
what length strip will be used?
3. What is the pH range of the IPG strip to be used?
4. How complex is the protein sample?
5. What staining or detection method will be used?
(for example, Bio-Safe™ Coomassie stain, silver stain,
etc.)
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