Anti-PAX8 antibody [EPR18715]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [EPR18715] (AB191870)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-OV-3 (Human ovarian cancer cell line) cells labeling PAX8 with ab191870 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nucleus staining on SK-OV-3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab191870 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [EPR18715] (AB191870)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH : OVCAR-3 (Human ovary adenocarcinoma cell line) cells labeling PAX8 with ab191870 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nucleus staining on NIH : OVCAR-3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab191870 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR18715] (AB191870)

Immunohistochemical analysis of paraffin-embedded Human endometrium carcinoma tissue labeling PAX8 with ab191870 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on tumor cells of the endometrium carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR18715] (AB191870)

Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling PAX8 with ab191870 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on tumor cells of thyroid carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-PAX8 antibody [EPR18715] (AB191870)

Immunohistochemistry (Frozen sections) analysis of mouse kidney tissue labelling PAX8 with ab191870 at a dilution of 1/500. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/1000). Nuclear staining is visible in the mouse kidney.

• WB

Supplier Data

Western blot - Anti-PAX8 antibody [EPR18715] (AB191870)

Blocking/Dilution buffer : 5% NFDM/TBST.

PAX8 is expressed in the excretory system and thyroid gland. (PMID : 1723950, 9590297, 1069301)

All lanes:

Western blot - Anti-PAX8 antibody [EPR18715] (ab191870) at 1/1000 dilution

Lane 1:

Human thyroid cancer lysate at 10 µg

Lane 2:

Human fetal liver lysate at 10 µg

Lane 3:

Human fetal heart lysate at 10 µg

Lane 4:

Human fetal kidney lysate at 10 µg

Lane 5:

Human fetal spleen lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PAX8 antibody [EPR18715] (AB191870)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PAX8 antibody [EPR18715] (ab191870) at 1/1000 dilution

Lane 1:

NIH:OVCAR-3 (Human ovary adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/10000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

false

Exposure time: 3min

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR18715] (AB191870)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 NIH : OVCAR-3 (Human ovary adenocarcinoma epithelial cell) cells and 5 µg of ab191870 [EPR18715]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods."

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR18715] (AB191870)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 NIH : OVCAR-3 (Human ovary adenocarcinoma epithelial cell) cells and 5 µg of ab191870 [EPR18715]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods."

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR18715] (AB191870)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 NIH : OVCAR-3 (Human ovary adenocarcinoma epithelial cell) cells and 5 µg of ab191870 [EPR18715]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods."