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Technical Data Sheet

CL9511

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C2C12 Calyculin A Control
Lysate
Price
Size

$75
100 μl

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Application
Dilution


WB
20 μl/lane



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Calyculin A is a serine/threonine phosphatase inhibitor that inhibits the activity of protein phosphatases PP1 and PP2A. Cells treated with calyculin A for 30 minutes can undergo significant threonine phosphorylation, as shown by western blotting using anti-Phosphoserine/threonine, cat.# PP2551, as compared to untreated, control cell lysates.

C2C12 is a mouse muscle myoblast cell line that is useful for studies of myoblast and osteoblast differentiation. Confluent cultures of C2C12 cells were serum starved for 2 hours. Cells were then either left untreated (Cat.# CL9511) or treated with Calyculin A (100 nM) for 30 minutes at 37°C (cat.# CL9521). Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 1 mM sodium fluoride in 10 mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.

Buffer and Storage
Cell Lysates are supplied at a concentration of 1 mg/ml in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. Stable for 1 year.
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