Connexin 43 Polyclonal Antibody

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	-Rabbit (1:2000)

Protocol tips
-Rabbit (1:2000)

Publication protocol

HCECs, MCECs, and isolated mitochondria from HCECs were lysed in lysate buffer (in mM: 25 HEPES, 150 NaCl, 10 MgCl2, and 1 EDTA, with 1% IGEPAL, 1% protease inhibitor, and 1% phosphate inhibitor, pH 7.5). Proteins were separated with a 10% SDS-polyacrylamide gel and transferred to nitrocellulose membranes. Blots were incubated with the following primary antibodies: anti-Cx40 (Santa Cruz, sc-20466; 1:2,000), anti-ATP5A (Santa Cruz, sc-136178; 1:2,000), anti-Actin (Santa Cruz, sc-1616; 1:4,000), anti-Cx37 (Thermo Fisher Scientific, 42-4400; 1:1,000), anti-Cx43 (Thermo Fisher Scientific, 71-0700; 1:2,000), anti-Cx45 (Thermo Fisher Scientific, 41-5800; 1:2,000), or anti-GAPDH (Thermo Fisher Scientific, MA5-15738; 1:4,000), followed by horseradish peroxidase-linked secondary antibody incubation. The immunoblots were identified with the SuperSignal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific, 34087). Band intensity was calculated with ImageJ 1.48v (National Institutes of Health), and the intensity data from focus proteins were normalized to GAPDH or Actin (for whole cell and cytosolic samples) or ATP5A (for mitochondrial samples) and are expressed in arbitrary units.

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