Beclin-1 (D40C5) Rabbit mAb

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Cells were lysed in RIPA buffer with PMSF (PMSF:RIPA=1:100) for 30 min on ice and then centrifuged for 15 min. Protein concentration of the supernatant was measured using a BCA protein assay kit.	Equal amounts of protein from cell extracts were separated by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred onto a PVDF (polyvinylidene fluoride) membrane (30). The membrane was incubated with primary antibodies overnight at 4°C and then with secondary antibodies conjugated with horseradish peroxidase.	Finally, the protein level was determined using a chemiluminescent approach in a dark room.

Upstream tips
Cells were lysed in RIPA buffer with PMSF (PMSF:RIPA=1:100) for 30 min on ice and then centrifuged for 15 min. Protein concentration of the supernatant was measured using a BCA protein assay kit.
Protocol tips
Equal amounts of protein from cell extracts were separated by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred onto a PVDF (polyvinylidene fluoride) membrane (30). The membrane was incubated with primary antibodies overnight at 4°C and then with secondary antibodies conjugated with horseradish peroxidase.
Downstream tips
Finally, the protein level was determined using a chemiluminescent approach in a dark room.

Publication protocol

Cells were lysed in RIPA buffer with PMSF (PMSF:RIPA=1:100) for 30 min on ice and then centrifuged for 15 min. Protein concentration of the supernatant was measured using a BCA protein assay kit. Equal amounts of protein from cell extracts were separated by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred onto a PVDF (polyvinylidene fluoride) membrane (30). The membrane was incubated with primary antibodies overnight at 4°C and then with secondary antibodies conjugated with horseradish peroxidase. Finally, the protein level was determined using a chemiluminescent approach in a dark room.

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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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