Anti-LC3 mAb

Autophagy assay cell type - C6

Experiment
Autophagy assay cell type - C6
Product
Anti-LC3 mAb from MBL international corporation
Manufacturer
MBL international corporation

Protocol tips

Upstream tips
- Storage buffer conditions:1 mg/mL in PBS/50% glycerol, pH 7.2

Publication protocol

C6 glioma cells were harvested and lysed in modified RIPA buffer (50 mM Tris-HCl, pH 7.4, 1% NP-40, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mM EDTA, 2 mM diisopropylfluorophosphate, 10 µg/ mL of leupeptin, 10 µg/mL of aprotinin, and 10 µg/mL of pepstatin). Cell lysates were centrifuged and proteins in the supernatants were electrophoresed in sodium dodecyl sulfate-polyacrylamide gels and transferred to Immobilon P membranes (Millipore, Billerica, MA). The membranes were blocked with 1% skim milk in 0.1% Tween 20/ PBS (PBS-T) and incubated with primary antibodies. Horseradish peroxidase-conjugated secondary antibodies were used and immunoreactive proteins were detected with ImmunoStar LD (Wako Pure Chemical Industries, Osaka, Japan).

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Manufacturer protocol

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