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Specifications

A431 cells express approximately 10e6 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.
Catalog #	A0001-52D
	Confluent cultures of A431 cells were serum starved overnight. Cells were then either left untreated or treated with human EGF (100 ng/ml) for 30 minutes at 37°C.
	Cells were lysed in 1% SDS, 1.0mM sodium ortho-vanadate, 10mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.
Applications	Suitable for use in SDS-PAGE/Western Blot .
Recommended Dilution	Western Blot: 20ul/well
	Optimal dilutions to be determined by the researcher
	Storage and Stability:
	For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Concentration	~1mg/ml
Form	Supplied in electrophoresis sample buffer (62.5mM Tris pH 7.2, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% b-mercaptoethanol).
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.