Hrp conjugated goat anti rabbit igg h l

Twenty micrograms of normal adult HB tissue lysate (Novus Biologicals) or heart tissue lysate (Novus Biologicals) was resolved on 4 to 20% gradient SDS-PAGE (Bio-Rad) and transferred to a polyvinylidene fluoride membrane (Millipore) for immunoblotting. Membranes were blocked with 5% BSA in TBST at RT for 1 h and then incubated with rabbit antisera (1:1000 dilution in 5% BSA/TBST) at RT for 1 h. Membranes were washed three times by TBST for 5 min each. Membranes were then incubated with HRP-conjugated goat anti-rabbit IgG (H + L) (1:4000 dilution; Southern Biotech) at RT for 1 h, and SuperSignal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific) was then added. The resulting chemiluminescence was recorded on a ChemiDoc XRS+ imaging system (Bio-Rad).

Total protein was extracted from skin tissues or from cultured cells and then subjected for Western blot as described previously 12 (link). The following antibodies and their corresponding dilutions were used: BRAP (Abcam, cat: ab181073; 1:2,000), IL-23 (Santa Cruz, cat: sc-271279,1:1,000;), IL-1β (Santa Cruz, cat: 12742,1:1,000), IL-10 (Santa Cruz, cat: 8438, 1:1,000), IL-17 (Affinity, cat: DF6127, 1:2,000), IL-17 (Santa Cruz, cat: sc-374218, 1:1,000), TNF-α (Santa Cruz, cat: sc-52746, 1:1,000), CK5 (Affinity, cat: AF5479, 1:2,000), β-Actin (Santa Cruz, cat: sc-47778, 1:5,000), CK14 (Santa Cruz, cat: sc-53253, 1:1,000), TGF-β1 (Abcam, cat: ab64715, 1:2,000), HRP-conjugated goat anti-Mouse IgG (H+L) (Proteintech, cat: SA00001-1, 1:5,000), HRP-conjugated goat anti-rabbit IgG(H+L) (SouthernBiotech, cat: 4050-05; 1: 5,000).