Catalog # AL9301

A431 + EGF (5 min) Treated

Lysate

Application Dilution
WB20 μl/lane

Size 100 μl

DATASHEET  




$75

A431 cells express approximately 106 epidermal growth factor (EGF) receptors at the cell surface. Upon stimulation with EGF, A431 cells exhibit a dramatic increase in phosphorylation of EGF receptors followed by activation of major cell signaling pathways, such as PKB/Akt and MAP kinase pathways. Downstream of these cell signaling pathways, a variety of cytoskeletal, cytoplasmic, and nuclear proteins become phosphorylated at different time points after EGF stimulation.

Confluent cultures of A431 cells were serum starved overnight. Cells were then either left untreated (Cat.# AL9201) or treated with human EGF (100 ng/ml) for 5 minutes at 37°C (Cat.# AL9301). 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.

Western blot analysis of A431 cells (20 µg/lane) serum starved overnight and treated with EGF (100 ng/ml) for 5 min. The blot was probed with anti-paxillin (PM1071) or anti-paxilin (Ser-178).



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.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains:

KIT SUMMARY