Anti-PAX6 antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody (AB238527)

Immunofluorescence staining of U-2 OS cells stained for PAX6 with ab238527 at 1/100 dilution. Nuclei are labeled with DAPI (Blue).

• WB

Supplier Data

Western blot - Anti-PAX6 antibody (AB238527)

Blocking buffer : 3% non-fat dry milk in TBST.

All lanes:

Western blot - Anti-PAX6 antibody (ab238527) at 1/1000 dilution

Lane 1:

SW480 (Human colorectal adenocarcinoma cell line) cell lysate at 25 µg

Lane 2:

22Rv1 (Human prostate carcinoma cell line) cell lysate at 25 µg

Lane 3:

NCI-H460 (Human lung cancer cell line) cell lysate at 25 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 25 µg

Lane 5:

Mouse brain tissue lysate at 25 µg

Lane 6:

Mouse eye tissue lysate at 25 µg

Lane 7:

Rat brain tissue lysate at 25 µg

Secondary

All lanes:

HRP Goat Anti-Rabbit IgG (H+L)

Predicted band size: 47 kDa

true

Exposure time: 30s

• WB

Lab

Western blot - Anti-PAX6 antibody (AB238527)

Western blot : Anti-PAX6 antibody (ab238527) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab238527 was shown to bind specifically to PAX6. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in PAX6 knockout cell line. To generate this image, wild-type and PAX6 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PAX6 antibody (ab238527) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PAX6 knockout A549 cell lysate at 20 µg

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 50 kDa

false