Bio-Rad ReadyPrep Protein Extraction Kit (Signal) - Manual (Page 12)

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
Sample calculation : If you precipitate 100 µg of protein and are going to
run 7 cm pH 3–10NL IPG strips (125 µl per strip) and silver stain the 2-D
gels, then you may want to solubilize the protein pellet in ~900 µl of
rehydration/sample buffer, which is enough to rehydrate about seven
7 cm IPG strips (~14 µg/strip). However, if you are planning to use a
24 cm pH 3–10NL IPG strip, then you may want to solubilize the protein
pellet in 410 µl of rehydration/sample buffer, which is enough to rehydrate
one 24 cm IPG strip (100 µg/strip). In this simple example, sample
complexity and IPG strip pH range were not addressed. As a general
rule, increased protein loads may be required for micro-range IPG strips
and for samples of higher protein complexity.
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