LC3B Antibody #2775

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Dilute primary Ab at 1:1000

Protocol tips
Dilute primary Ab at 1:1000

Publication protocol

Cells and tissues were lysed in RIPA buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% sodium deoxycholate, 0.1% SDS, and 1% triton X-100 supplemented with phosphatase and protease inhibitor cocktail) (Roche). Lysates were centrifuged at 15,000 g for 15 min at 4°C, and the supernatant was collected. Protein concentration was measured using a BCA protein assay kit (Thermo Fisher Scientific). Lysates were boiled in Laemmli sample buffer for 5 min, separated by SDS-PAGE, transferred to polyvinylidene fluoride membranes, and probed with primary antibodies (1:2000). After incubation with secondary antibodies conjugated with horseradish peroxidase (1:2000; Cell Signaling), chemiluminescence was detected using LAS1000 systems (Fujifilm, Tokyo, Japan). Immunoblot densitometric quantification was performed using ImageJ (NIH).

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for LC3B Antibody #2775 below.

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Videos

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