Manual

Preparation for IEF on an IPG Strip
Following step 17, the sample extract can be loaded onto
an IPG strip after appropriate dilution. See Table 3 for
recommendations on how much protein sample to load
onto an IPG strip. Dilution of the sample can be done using
protein solubilization buffer (PSB) as a rehydration/sample
buffer. However, some critical components need to be
added to the PSB solution to make it IPG-compatible
(Table 2).
The suggestions made in Table 3 are a general rule of
thumb. Increased protein loads may be required for
micro-range IPG strips and for samples of higher protein
complexity.
Table 3. Recommended protein loads for IPG
strips.
IIPPGG SSttrriipp LLeennggtthh
77 ccmm 1111 ccmm 1177 ccmm 1188 ccmm 2244 ccmm
Rehydration volume/strip 125 µl 185 µl 300 µl 315 µl 410 µl
Protein load
Silver stain 5–20 µg 20–50 µg 50–80 µg 50–80 µg 80–150 µg
Coomassie G-250 50–100 µg 100–200 µg 200–400 µg 200–400 µg 400–800 µg
Flamingo
™
,
SYPRO Ruby 2.5–75 µg 10–150 µg 25–300 µg 25–300 µg 40–600 µg
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