Beclin-1 (D40C5) Rabbit mAb

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Beclin-1 (D40C5) Rabbit mAb detects endogenous levels of total Beclin-1 protein."	Dilute primary Ab at 1:1000 and incubate overnight at 4℃ and for IF at 1:200

Upstream tips
Beclin-1 (D40C5) Rabbit mAb detects endogenous levels of total Beclin-1 protein."
Protocol tips
Dilute primary Ab at 1:1000 and incubate overnight at 4℃ and for IF at 1:200

Publication protocol

Cells were lysed with RIPA lysis buffer containing phosphatase inhibitor cocktail and protease inhibitor cocktail. Lysed cells were collected and centrifuged at 13,000 rpm for 10 min. The supernatants were transferred to 1.5 ml e-tube as whole cell lysates. The protein concentration in whole cell lysates were measured via the BCA method. An equal amount of proteins was subjected by SDS-polyacrylamide gel electrophoresis and transferred to the PVDF membrane. The PVDF membranes were blocked with 5% skim milk or 1% bovine serum albumin (BSA) in Tris-buffered saline-Tween 20 (TBS-T). Then, an antibody reaction for the primary antibodies diluted by 5% skim milk or 1% BSA solution was performed overnight at 4℃, and then the PVDF membrane was washed three times with TBS-T for 5 min each time. Next, secondary antibodies conjugated HRP were probed onto the PVDF membrane for 1 h at room temperature, and chemiluminescent substrate reaction for HRP was performed after washing the membrane with TBS-T. The protein band for each target was imaged and photographed by Luminescent Image Analyzer LAS-4000 (Fujifilm, Tokyo, Japan). Protein bands were densitometrically analyzed using Scion Image (Scion Corporation, Frederick, MD, 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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