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Technical Data Sheet

AL9201

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A431 EGF Control
Lysate
Price
Size

$75
100 μl

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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).

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
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.

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.
imageRelated Products
AL9001
A431 Calyculin A Control Lysate

AL9101
A431 + Calyculin A (30min) Lysate

AL9301
A431 + EGF (5 min) Lysate

AL9401
A431 Pervanadate Control Lysate

AL9501
A431 + Pervanadate Lysate

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