VDAC1/Porin Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000)

Protocol tips
-Mouse (1:1000)

Publication protocol

BMDMs were washed in ice-cold PBS, and 500 µl of ice-cold NP-40 (50 mM Tris, pH 7.8, 50 mM NaCl, 1% (vol/vol) Nonidet-P40, 5 mM EDTA, and 10% (vol/vol) glycerol) was added. Cells were dissolved on the shaker for 30 min at 4 °C and then centrifuged at 330×g for 10 min at 4 °C. The pellets were washed twice in 1 ml of ice-cold PBS and resuspended in 500 µl of ice-cold PBS. 2 mM disuccinimydylsuberate (Sangon biotech, C100015) was added to the resuspended pellets, which were incubated for 30 min with rotation at room temperature. Samples were then centrifuged at 330×g for 10 min at 4 °C. The supernatants were removed, and the cross-linked pellets were resuspended in 30 µl of sample buffer. Samples were boiled for 10 min at 101 °C and analyzed by western blotting.

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Manufacturer protocol

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