Atg5 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Lyse cells in RIPA buffer (50 mM Tris, 1.0 mM EDTA, 150 mM NaCl, 0.1% Triton X-100, 1% sodium deoxycholate, 1 mM PMSF)	-Dilute primary Ab at 1:1000 and at 4 °C overnight,

Upstream tips
- Lyse cells in RIPA buffer (50 mM Tris, 1.0 mM EDTA, 150 mM NaCl, 0.1% Triton X-100, 1% sodium deoxycholate, 1 mM PMSF)
Protocol tips
-Dilute primary Ab at 1:1000 and at 4 °C overnight,

Publication protocol

Cells were lysed with RIPA buffer (50 mM Tris, 1.0 mM EDTA, 150 mM NaCl, 0.1% Triton X-100, 1% sodium deoxycholate, 1 mM PMSF). Protein concentrations were quantified by the BCA protein assay kit (Pierce, 23225). Proteins were resolved on 12% SDS-PAGE, then transferred to PVDF (Bio-Rad) membranes. After blocking, the membranes were incubated with primary antibodies at 4 °C overnight, then incubated with secondary antibodies at room temperature for 1 h. Target proteins were examined using Enhanced Chemiluminescence reagents (Millipore, WBKLS0100).

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Atg5 Antibody below.

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