Anti-CYP1B1 antibody [EPR14972] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP1B1 antibody [EPR14972] - BSA and Azide free (AB232645)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185954).

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

• WB

Lab

Western blot - Anti-CYP1B1 antibody [EPR14972] - BSA and Azide free (AB232645)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

All lanes:

Western blot - Anti-CYP1B1 antibody [EPR14972] - C-terminal (<a href='/en-us/products/primary-antibodies/cyp1b1-antibody-epr14972-c-terminal-ab185954'>ab185954</a>) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CYP1B1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CYP1B1 knockout A549 cell lysate (<a href='/en-us/products/cell-lysates/human-cyp1b1-knockout-a549-cell-lysate-ab269641'>ab269641</a>)

Lane 2:

Western blot - Human CYP1B1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-cyp1b1-knockout-a549-cell-line-ab269476'>ab269476</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Lane 5:

SiHa cell lysate at 20 µg

Lane 6:

Human Brain cell lysate at 20 µg

Lane 7:

Human Kidney cell lysate at 20 µg

Lane 8:

Human Liver cell lysate at 20 µg

Lane 9:

HepG2 cell lysate at 20 µg

Lane 10:

LoVo cell lysate at 20 µg

Secondary

All lanes:

HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 61 kDa

Observed band size: 55 kDa

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