Bio-Rad MicroRotofor™ Cell Lysis Kits - Manual (Page 12)

Table 3. Recommended protein loads for IPG
strips.
IIP PG G S Sttrriip p L Leen ng gtth h
7 7 ccm m 1 11 1 ccm m 1 17 7 ccm m 1 18 8 ccm m 2 24 4 ccm m
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
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.
Lyticase Enzyme
This enzyme is prepared from Arthrobacter luteus. The
primary yeast lytic activity is beta1, 3-glucan
laminaripentaohydrolase, which hydrolyzes glucose
polymers at the beta-1,3-glucan linkages, releasing
laminaripentaose as the principal product.
Unit definition: One lytic unit is defined as a 10% decrease
in absorbance at A
800
in 30 min.
Assay condition: 50 mM potassium phosphate, pH 7.5,
10 mM
β-mercaptoethanol in 1 ml yeast cell suspension of
A
800
0.8 to 1.0.
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