Anti-PAX8 antibody [1F8-3A8]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [1F8-3A8] (AB183573)

Immunofluorescent analysis of PAX8 (green) in 293T (left panel) and COS7 cells (right panel) using ab183573 at a 1/50 dilution. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% blocker BSA for 15 minutes at room temperature. A DyLight 488-conjugated goat anti-mouse IgG secondary antibody was used at a dilution of 1/400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [1F8-3A8] (AB183573)

Immunohistochemistry analysis of PAX8 showing staining in the nucleus of paraffin-embedded human kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab183573 diluted in 3% BSA-PBS at a dilution of 1 : 100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

• WB

Supplier Data

Western blot - Anti-PAX8 antibody [1F8-3A8] (AB183573)

4-20% Tris-HCl polyacrylamide gel. Blocked with 5% BSA/TBST for at least 1 hour.

All lanes:

Western blot - Anti-PAX8 antibody [1F8-3A8] (ab183573) at 1/1000 dilution

Lane 1:

SKOV-3 cell lysate at 60 µg

Lane 2:

COS7 cell lysate at 60 µg

Lane 3:

Negative control HeLa whole cell lysates at 60 µg

Secondary

All lanes:

HRP-conjugated goat anti-mouse IgG at 1/500 dilution

Predicted band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-PAX8 antibody [1F8-3A8] (AB183573)

4-20% Tris-HCl polyacrylamide gel. Blocked with 5% BSA/TBST for at least 1 hour.

All lanes:

Western blot - Anti-PAX8 antibody [1F8-3A8] (ab183573) at 1/1000 dilution

Lane 1:

untransfected HEK293T cell lysate at 25 µg

Lane 2:

HEK293T cell lysate over expressing PAX8 at 25 µg

Secondary

All lanes:

HRP-conjugated goat anti-mouse IgG at 1/500 dilution

Predicted band size: 48 kDa

false