Owner manual

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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