Cyclophilin B Antibodies

Immunofluorescent analysis of Cyclophilin B. Cyclophilin B (green) staining showing staining in the in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclophilin B polyclonal antibody (Cat. No. PA1-027A) in 3% BSA-PBS at a dilution of 1:500 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), IMR-32 (Lane 2), U-87 MG (Lane 3), PC-3 (Lane 4), HeLa (Lane 5), tissue extracts of Mouse Liver (Lane 6) and Mouse Heart (Lane 7). The blot was probed with Anti-Cyclophilin B Rabbit polyclonal Antibody (Cat. No. TAB1002, 0.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Cat. No. A27036, 0.25 µg/mL, 1:4000 dilution). A 23 kDa band corresponding to Cyclophilin B was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex NuPAGE12 % Bis-Tris gel (Cat. No. NP0342BOX), XCell SureLock Electrophoresis System (Cat. No. EI0002) and Novex Sharp Pre-Stained Protein Standard (Cat. No. LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot 2 Dry Blotting System (Cat. No. IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind Flex Western Starter Kit (Cat. No. SLF2000S). Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Cat. No. 32106).