Recombinant Anti-CYP1B1 antibody [EPR14972] - C-terminal (ab185954)

Western blot: Anti-CYP1B1 antibody [EPR14972] (ab185954) staining at 1/5000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab185954 was shown to bind specifically to CYP1B1. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in CYP1B1 knockout cell line ab269476 (knockout cell lysate ab269641). To generate this image, wild-type and CYP1B1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 8 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Immunohistochemical analysis of paraffin-embedded Human renal adenocarcinoma tissue labeling CYP1B1 with ab185954 at 1/500 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.