A431 Cells, EGF-Stimulated

Biologicals / Biologicals-Cells-Extracts, Lysates

-20°C

A431 cells were cultured for 20 minutes in the presence of 50ng/ml EGF. Cells were lysed in modified RIPA buffer (50mM Tris-HCl, pH 7.4, 1% NP40, 0.25% sodium deoxycholate, 150mM NaCl, 1mM EDTA, 1mM PMSF, 1ug/ml aprotinin, 1ug/ml leupeptin, 1ug/ml pepstatin, 1mM Na3VO4, 1mM NaF) and diluted with non-reducing sample buffer (31mM Tris-HCl, pH 6.8, 5% glycerol, 1% SDS, 0.002% bromophenol blue).

Cellular protein preparation from A431 cells containing phosphorylated proteins, including the EGF receptor.

Recommended Dilutions:
Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for immunoblot analysis. This preparation may be used as a positive control for antibodies.

Storage and Stability:
For long-term storage, aliquot and store at -20°C. Aliquots are stable for 6 months after receipt at -20°C after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.