Anti-MAGEA4 antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAGEA4 antibody (AB229011)

Paraffin-embedded human breast cancer tissue stained for MAGEA4 using ab229011 at 1/500 dilution in immunohistochemical analysis.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MAGEA4 antibody (AB229011)

4% paraformaldehyde-fixed A-375 (human malignant melanoma cell line) cells stained for MAGEA4 (green) using ab229011 at 1/500 dilution in ICC/IF.

Nuclear counterstain : Hoechst 33342 (blue).

• WB

Supplier Data

Western blot - Anti-MAGEA4 antibody (AB229011)

12% SDS-PAGE gel.

All lanes:

Western blot - Anti-MAGEA4 antibody (ab229011) at 1/2000 dilution

All lanes:

A549 (human lung carcinoma cell line) whole cell lysate at 30 µg

Predicted band size: 35 kDa

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• WB

Lab

Western blot - Anti-MAGEA4 antibody (AB229011)

False colour image of Western blot : Anti-MAGEA4 antibody staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab229011 was shown to bind specifically to MAGEA4. A band was observed at 40 kDa in wild-type A431 cell lysates with no signal observed at this size in MAGEA4 knockout cell line ab273857 (knockout cell lysate ab273811). To generate this image, wild-type and MAGEA4 knockout A431 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:

SH-SY5Y cell lysate at 1/500 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human MAGEA4 knockout A-431 cell lysate (ab273811) at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

Western blot 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: 40 kDa

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