ICAM-1 Antibody (G-5): sc-8439

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Western blot analysis was performed as previously described [9]. For coimmunoprecipitation analysis, cells were lysed on ice in modified RIPA buffer (50mM Tris pH 7.4 150 mM NaCl, 1%NP-40, 0.1% SDS, 0.5% SD Dexoxycholate, 5mM NaF, 2.5 mM Nappi, and protease inhibitor (#04693159001, Roche) for 30 minutes and isolated by centrifugation (15 min, 10000g, 4°C). Supernatants were precleared during 1 hour at 4°C with Magna CHIP protein G bead (#16-662, Millipore) and normal mouse IgG (#sc-2025, Santa Cruz technology) and the cleared lysate incubated with primary antibody or IgG overnight at 4°C. Immune complexes were captured by adding 35μl of protein G magnetic beads, rotated for 1h at 4°C and washed three times with lysis buffer (without SDS, Sodium Deoxycholate and protease inhibitors). Immunoprecipitation products were separated by SDS-PAGE.

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Manufacturer protocol

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