cytochrome c Antibody (A-8): sc-13156

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

For Western blot analysis, protein samples were separated by SDS-PAGE and then transferred to polyvinylidene difluoride membranes. Membranes were blocked with 5% nonfat dry milk in PBS, and proteins were detected by incubation with primary antibodies diluted in PBS followed by incubation with secondary antibodies (raised in goat against the appropriate species) conjugated to horseradish peroxidase (HRP) and diluted in PBS unless otherwise stated. Antibody binding was detected with an enhanced chemiluminescence (ECL) kit (Pierce) and film. The following antibodies were used in Western blots: rat monoclonal anti-HA antibody directly conjugated to HRP (Roche, catalog no. ab 3F10 [diluted 1:1,000]), rabbit polyclonal anti-MIC60 (Proteintech, catalog no. 10179-1-AP [diluted 1:1,000]), rabbit monoclonal anti-SAM50 (Abcam, Inc., catalog no. AB167430 [diluted 1:5,000]), mouse polyclonal anti-MIC19 (Abcam, Inc., catalog no. AB69328 [diluted 1:1,000]), rabbit monoclonal anti-EEA1 (Cell Signaling, catalog no. 3288 [diluted 1:1,000]), rabbit polyclonal anti-TOM20 (Santa Cruz Biotechnology, catalog no. sc-11415 [diluted 1:1,000]), rabbit polyclonal anti-TOM40 (Santa Cruz Biotechnology, catalog no. sc-11414 [diluted 1:500]), mouse monoclonal anti-cytochrome c (Santa Cruz Biotechnology, catalog no. sc-13156 [diluted 1:1,000]), mouse monoclonal anti-GAPDH (Calbiochem, catalog no. CB-1001 [diluted 1:10,000]), mouse monoclonal anti-DRP-1 (Abcam, Inc., catalog no. AB56788 [diluted 1:1,000]), mouse anti-metaxin1 (Santa Cruz Biotech, catalog no. sc-514469 [diluted 1:1,000]), anti-ATP5A (Abcam, Inc., catalog no. AB14748 [diluted 1:1,000]), rabbit polyclonal anti-SAG1 (generated previously to recombinant SAG1 [diluted 1:20,000]), rabbit anti-GRA7 (generated previously [50] [diluted 1:10,000]), and rabbit polyclonal anti-MAF1b (generated in this study as described below [diluted 1:10,000 in PBS plus 5% milk]).

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Manufacturer protocol

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