Anti-HE4 antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HE4 antibody (AB273130)

Paraffin-embedded human cervix (uterine) tissue stained for HE4 using ab273130 at 1/200 dilution in immunohistochemical analysis.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HE4 antibody (AB273130)

PFA-fixed, Triton X-100-permeabilized A431 (Human epidermoid carcinoma cell line) cells stained for HE4 using ab273130 at 2 μg/ml (green) in ICC/IF.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HE4 antibody (AB273130)

Paraffin-embedded human ovary tissue stained for HE4 using ab273130 at 1/50 dilution in immunohistochemical analysis.

Shows low expression as expected.

Retrieval method : HIER pH6.

• WB

Lab

Western blot - Anti-HE4 antibody (AB273130)

False colour image of Western blot : Anti-HE4 antibody staining at 0.04 ug/ml, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab273130 was shown to bind specifically to HE4. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 Optiblot (ECL reagent ab133456) and imaged with 4 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-HE4 antibody (ab273130) at 0.04 µg/mL

Lane 1:

OVCAR-3 cell lysate at 20 µg

Lane 2:

K562 cell lysate at 20 µg

Predicted band size: 13 kDa

Observed band size: 20 kDa

false

• WB

Lab

Western blot - Anti-HE4 antibody (AB273130)

False colour image of Western blot : Anti-HE4 antibody staining at 0.4 ug/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab273130 was shown to bind specifically to HE4. A band was observed at 20 kDa in treated wild-type OVCAR-3 cell lysates with no signal observed at this size in WFDC2 knockout cell line ab275410 (knockout cell lysate ab275536). To generate this image, wild-type and WFDC2 knockout OVCAR-3 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-HE4 antibody (ab273130) at 0.4 µg/mL

Lane 1:

Wild-type OVCAR-3 Treated Brefeldin A (<a href='/en-us/products/sample-preparation-kits/brefeldin-a-solution-1000x-ab193369'>ab193369</a>) (5 ug/mL, 4 h), Monensin (<a href='/en-us/products/sample-preparation-kits/monensin-solution-1000x-ab193381'>ab193381</a>) (2 uM, 4 h) cell lysate at 20 µg

Lane 2:

Wild-type OVCAR-3 Vehicle Control Brefeldin A (<a href='/en-us/products/sample-preparation-kits/brefeldin-a-solution-1000x-ab193369'>ab193369</a>) (0 ug/mL, 4 h), Monensin (<a href='/en-us/products/sample-preparation-kits/monensin-solution-1000x-ab193381'>ab193381</a>) (0 uM, 4 h) cell lysate at 20 µg

Lane 3:

WFDC2 knockout OVCAR-3 Treated Brefeldin A (<a href='/en-us/products/sample-preparation-kits/brefeldin-a-solution-1000x-ab193369'>ab193369</a>) (5 ug/mL, 4 h), Monensin (<a href='/en-us/products/sample-preparation-kits/monensin-solution-1000x-ab193381'>ab193381</a>) (2 uM, 4 h) cell lysate at 20 µg

Lane 4:

WFDC2 knockout OVCAR-3 Vehicle Control Brefeldin A (<a href='/en-us/products/sample-preparation-kits/brefeldin-a-solution-1000x-ab193369'>ab193369</a>) (0 ug/mL, 4 h), Monensin (<a href='/en-us/products/sample-preparation-kits/monensin-solution-1000x-ab193381'>ab193381</a>) (0 uM, 4 h) cell lysate at 20 µg

Observed band size: 20 kDa

false

• WB

Supplier Data

Western blot - Anti-HE4 antibody (AB273130)

All lanes:

Western blot - Anti-HE4 antibody (ab273130) at 0.4 µg/mL

All lanes:

EFO-21 (Human cell line) cell lysate

Predicted band size: 13 kDa

false