BECN1 Antibody (H-300)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Dilute Primary Ab -1:200. - Incubate Ab at overnight at 4 °C

Protocol tips
- Dilute Primary Ab -1:200. - Incubate Ab at overnight at 4 °C

Publication protocol

Total proteins were extracted from GFP-LC3/CCL149 cells and rat lung tissues in lysis buffer. The protein concentrations were determined using a Pierce Bicinchoninic Acid Protein Assay kit (Biyuntian, Shanghai, China, #P0010). Fifty micrograms of protein was subjected to SDS–polyacrylamide gel electrophoresis (12%PAGE; Amresco) and transferred to a polyvinylidene fluoride membrane (Millipore) followed by incubation overnight at 4 °C with the following primary antibodies: LC3 (Abcam, Cambridge, United Kingdom, #ab62341; 1:200 dilution), Beclin-1 (Santa Cruz, CA, USA, #sc-11427; 1:200 dilution), NF-κB (Bioworld, CA, USA, #BS1257; 1:600 dilution), and IκB-α (Bioworld, CA, USA, #BS3601; 1:600 dilution). After incubation with peroxidase-conjugated secondary antibodies (BA1060, at 1:50,000 dilution), the bands were visualized using Bio-Rad Chemi DocTM XRS+ (Bio-Rad). Protein expression levels were normalized to the corresponding β-actin levels.

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Manufacturer protocol

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