Beclin-1 (D40C5) Rabbit mAb

Autophagy assay cell type - HepG2

Experiment
Autophagy assay cell type - HepG2
Product
Beclin-1 (D40C5) Rabbit mAb from Cell Signaling Technology
Manufacturer
Cell Signaling Technology

Protocol tips

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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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - HepG2 using Beclin-1 (D40C5) Rabbit mAb from Cell Signaling Technology.

Paper title
SPHK1 (sphingosine kinase 1) induces epithelial-mesenchymal transition by promoting the autophagy-linked lysosomal degradation of CDH1/E-cadherin in hepatoma cells
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

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