HER2/ErbB2 (29D8) Rabbit mAb #2165

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Rabbit (1:1000)

Protocol tips
-Rabbit (1:1000)

Publication protocol

Tissue samples were first minced and then suspended in cell lysis buffer containing 20 mM Tris-HCl, pH7.2, 150 mM NaCl, 50 mM NaF, 1 mM EDTA, 10% glycerol, 1% triton-X-100, 1 mM sodium orthovanadate and a protease inhibitor cocktail. The suspensions were sonicated on ice, incubated at 4oC for 4 hours with rocking, centrifuged at 12,000 rpm for 10 minutes, and used for immunoblotting analyses. Cell lysates were prepared in the same buffer without vigorous sonication. Electrophoretic and immunoblotting analyses were conducted as described previously (21, 38). Specific antibodies were used one-after-another for detection of different proteins on the same membrane. For this, some full-length blots contain bands for different proteins (e.g. pAkt and pERK1/2). X-ray films were digitalized by scanning at 600 dpi resolution using Adobe Photoshop software.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for HER2/ErbB2 (29D8) Rabbit mAb #2165 below.

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