LC3B (D11) XP® Rabbit mAb (Biotinylated)

Protocol tips

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	- Incubate with Primary Ab dilution-1:1000 at overnight at 4°C

Protocol tips
- Incubate with Primary Ab dilution-1:1000 at overnight at 4°C

Publication protocol

Whole cell lysates was prepared by addition of lysis buffer (50mM Tris PH 7.4, 150mM NaCl, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate). 10μg of protein lysate was separated by SDS-PAGE and then transferred to polyvinylidene difluoride membranes. After blocking with 5% non-fat milk, the membranes were incubated with primary antibodies, β-actin (1:5000; Chemicon, Temecula, CA), Bcl-2 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA), Bcl-XL (1:1000; Cell Signaling Technology, Beverly, MA), Bax (1:1000; Cell Signaling Technology, Beverly, MA), Cleaved Caspase 3 (1:1000; Cell Signaling Technology, Beverly, MA), LC3 (1:1000; Cell Signaling Technology, Beverly, MA), P-p70S6K (1:1000; Cell Signaling Technology, Beverly, MA), p70S6K (1:1000; Cell Signaling Technology, Beverly, MA), p-AMPK (1:1000; Cell Signaling Technology, Beverly, MA), AMPK (1:1000; Cell Signaling Technology, Beverly, MA), p-mTOR (1:1000; Cell Signaling Technology, Beverly, MA), mTOR (1:1000; Cell Signaling Technology, Beverly, MA), p-ULK1(Ser757) (1:1000; Cell Signaling Technology, Beverly, MA) overnight at 4°C. After incubation with secondary antibody, signals were detected by ECL (Amersham Pharmacia Biotech, Arlington Height, IL) on a X-ray film, which was scanned and quantified using Image J software (National Institute of Mental Health Bethesda). Five independent experiments with duplicate samples were performed.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for LC3B (D11) XP® Rabbit mAb (Biotinylated) below.

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