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Connexin 26 / GJB2 Antibody (710500) in WB
Western blot analysis of Connexin26 (Cx26) was performed by loading 30 ug of protein from mouse liver homogenate (not boiled) and 7 ul of EZ-RUN Prestained Rec Protein Ladder (Product # BP3603-500) per well onto a 10% Tris-Hcl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked in blocking buffer composed of 20 mM Tris-HCl pH 8, 150 mM NaCl and 0.2% Tween-20 (TBST) supplemented with 5% skim milk powder (Carnation) for 1 hr at room temperature. Cx26 was detected at ~26 kDa using an anti-Cx26 antibody (Product # 71-0500, lane 2) at a dilution of 2 ug/ml in TBST supplemented with 1% skim milk powder and incubated overnight at 4°C on a rocking platform, followed by washing and incubation with a goat anti-rabbit IgG horse radish peroxidase (HRP) conjugated secondary antibody at a dilution of 1:5,000 in TBST for 1 hr at room temperature. Chemiluminescent detection was performed using Pierce ECL Western blotting substrate (Product # 32209). Data courtesy of Dr. James Nagy's lab.
