Phospho-mTOR (Ser2448) Antibody

Autophagy assay cell type - HEK 293

Experiment
Autophagy assay cell type - HEK 293
Product
Phospho-mTOR (Ser2448) Antibody from Cell Signaling Technology
Manufacturer
Cell Signaling Technology

Protocol tips

Protocol tips
Dilute primary Ab at 1:1000

Publication protocol

Bacterial whole cell lysates and infected or uninfected host cells were lysed in a sample buffer containing 200 mM DTT, 8% SDS and 5% 2-mercaptoethanol. The procedures used for SDS-PAGE and immunoblot analysis were performed as described previously (Laemmli, 1970). The following primary antibodies were used: anti-GFP (Roche, 1:5,000); anti-LC3B (#2775, CST); anti-p-mTOR (Ser2448; #2971, CST); anti-p-p70S6K (Thr389; #9206, CST); anti-4E-BP1 (#9644, CST); anti-p-4E-BP1 (Thr37/46; #2855, CST); anti-p-AMPK (Thr172; #2535, CST); anti-p-eIF2α (Ser51; #3398, CST), anti-α-actin (Sigma-Aldrich, 1:10,000); anti-LLO (Abcam, 1:4,000); and anti-VCC (1:10,000; Ou et al., 2009). Secondary antibodies, horseradish peroxidase conjugated goat anti-rabbit (Agrisera) and rabbit anti-mouse (Dako) were used at 1:20,000 dilutions. The intensity of immunoblot reaction bands were analyzed semi-quantitatively using Quantity One 1-D analysis software (Bio-Rad) or Image J.

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Papers

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Paper title
A Novel Role of Membrane Vesicles in Inhibition of Autophagy and Cell Death
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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Phospho-mTOR (Ser2448) Antibody below.

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