Bio-Rad ReadyPrep™ Protein Extraction Kit (Membrane II) - Manual (Page 16)

Section 6
Appendix
2-D Rehydration/Sample Buffer Volume
Before IEF and 2D gel electrophoresis, the protein sample
may need to be diluted to achieve the desired protein load
for the chosen stain. To best determine the volume of
diluent to use, consider the IPG strip length, the pH
gradient of the IPG strip, and the staining or detection
method. 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.
IPG strip length 7 cm 11 cm 17 cm 18 cm 24 cm
Rehydration 125 µl 185 µl 300 µl 315 µl 410 µl
volume per strip
Protein load– 5–20 µg 20–50 µg 50–80 µg 50–80 µg 80–150 µg
Silver stain
Protein load– 50–100 µg 100–200 µg 200–400 µg 200–400 µg 400–800 µg
Coomassie G-250
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