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Technical Data Sheet

CL8041

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C2C12 + Pervanadate
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
Pervanadate is a protein tyrosine phosphatase inhibitor that is commonly used to increase tyrosine phosphorylation in cells. When cells are treated with pervanadate for 30 minutes they undergo significant tyrosine phosphorylation, as shown by western blotting using anti-Phosphotyrosine.

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 hrs, and then either left untreated (Cat.# CL8031) or treated with pervanadate (1 mM) for 30 minutes at 37°C (Cat.# CL8041). Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 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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