Anti-Beclin 1 antibody (ab62557)

Protocol tips

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	- Primary antibody dilution:1:2000

Protocol tips
- Primary antibody dilution:1:2000

Publication protocol

Cells were lysed on ice for 15 min in 16RIPA lysis buffer. Protein concentration was measured using the Bradford assay.
The amount of protein was approximately 30,60 mg (concentration measured by Bradford method: 1.5,3 mg/ml) per sample. Equal amounts of cell extracts were boiled for 5 min in the presence of 1% 2-mercaptoethanol and 2% sodium dodecyl sulphate (SDS). Samples were electrophoresed on 15% SDS-poly-acrylamide gels, and then transferred to polyvinylidene difluoride (PVDF) membrane. Membranes were blocked with 5% (wt/vol) milk proteins/Tris-buffered saline and incubated overnight at 4uC with the primary antibody: rabbit anti-LC3 (2 mg/ml), rabbit antiBeclin 1 (1 mg/ml), rabbit anti-Atg7 (1 mg/ml), rabbit anti-cleaved caspase 3 (2 mg/ml), anti-phospho-P44/42 MAP Kinase (2 mg/ ml), anti-P44/42 MAP Kinase (2 mg/ml), anti-SAPK/JNK (2 mg/
ml), anti-phospho-SAPK/JNK (2 mg/ml), anti-p38MAP Kinase (2 mg/ml) and anti-phospho-p38 MAP Kinase (2 mg/ml). Bound antibodies (dilution:1:2000) were detected with appropriate


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Manufacturer protocol

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