GC Antibody (Center)

Western blot analysis of GC Antibody (Center) in mouse lung tissue lysates (35 ug/lane). GC (arrow) was detected using the purified Pab.

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue was performed on the Leica BOND RXm. Samples were incubated with primary antibody (1/500) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.

All lanes: Anti-GC Antibody (Center) at 1:2000 dilution. Lane 1: 293T/17 whole cell lysate. Lane 2: Hela whole cell lysate. Lane 3: LNCaP whole cell lysate. Lane 4: MCF-7 whole cell lysate. Lane 5: U-2OS whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 60 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Overlay histogram showing Hela cells stained (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight 488 Conjugated Highly Cross-Adsorbed (1583138) at 1/200 dilution for 40 min at 37°C. Isotype control antibody (blue line) was rabbit IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 events was performed.