Beclin-1 (D40C5) Rabbit mAb

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	For WB, dilute primary Ab at 1:1000 and for immunoprecipitation at 1:100. For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight

Protocol tips
For WB, dilute primary Ab at 1:1000 and for immunoprecipitation at 1:100. For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight

Publication protocol

Cells were washed 3 times with phosphate-buffered saline (Invitrogen, 10010023) and then harvested. The cell lysates were extracted in co-immunoprecipitation buffer.16 As described previously,16,33 total cell lysates (5 mg of protein) were subjected to immunoprecipitation with the indicated antibodies overnight at 4°C and then incubated with protein A/G Plus-agarose (Santa Cruz Biotechnology, sc-2003) for 24 h at 4°C. After being washed 3 times, the immunocomplexes were mixed with 2×SDS loading buffer and boiled for 5 min. For the western blot analysis, co-precipitates or total cell lysates were electrophoresed by SDS-PAGE and transferred to PVDF membranes (Millipore, ISEQ00010). The membranes were immunoblotted with the appropriate antibodies and then developed using a ChemiImager 5500 imaging system (α Innotech Co., USA).

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Beclin-1 (D40C5) Rabbit mAb below.

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