Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human thyroid tissue staining PAX8 with ab239363 at a 1/2000 dilution (0.247 μg/ml), ab133322 anti-Thyroid Peroxidase used at a 1/2000 dilution (0.377 μg/ml) and ab151539 anti-Thyroglobulin used at a 1/6000 dilution (0.14 μg/ml).

Panel A : merged staining of anti-PAX8 (green; Opal^TM520), anti-Thyroid Peroxidase (magenta; Opal^TM690) and anti-Thyroglobulin (gray; Opal^TM570) on human thyroid.

Panel B : anti-PAX8 staining nucleus of follicular epithelial cells in human thyroid.

Panel C : anti-Thyroid Peroxidase staining cytoplasm of follicular epithelial cells in human thyroid.

Panel D : anti-Thyroglobulin staining colloid in human thyroid.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab239363, ab133322 and ab151539 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^TM 4-color kit using Opal Polymer HRP Ms + Rb secondary reagent. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling PAX8 with ab239363 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human ovarian cancer (PMID : 21317881). The section was incubated with ab239363 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human thyroid tissue labeling PAX8 with ab239363 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human thyroid (PMID : 21317881). The section was incubated with ab239363 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH : OVCAR-3 (human ovary adenocarcinoma epithelial cell) cells labelling PAX8 with ab239363 at 1/500 dilution (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH : OVCAR-3 (human ovary adenocarcinoma epithelial cell) cells labelling PAX8 with ab239363 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (Green). Confocal image showing nuclear staining in NIH : OVCAR-3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-OV-3 (human ovarian cancer epithelial cell) cells labelling PAX8 with ab239363 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in SK-OV-3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IP

Unknown

Immunoprecipitation - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

PAX8 was immunoprecipitated from 0.35 mg SK-OV-3 (human ovarian cancer epithelial cell), whole cell lysate 10 ug with ab239363 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab239363 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : SK-OV-3 (human ovarian cancer epithelial cell), whole cell lysate 10 ug

Lane 2 : ab239363 IP in SK-OV-3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab239363 in SK-OV-3 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 49 seconds

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-PAX8 antibody [EPR23508-20] (<a href='/ko/products/primary-antibodies/pax8-antibody-epr23508-20-ab239363'>ab239363</a>)

Predicted band size: 48 kDa

false

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse kidney tissue labeling PAX8 with ab239363 at 1/50 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Nuclear staining on mouse kidney. is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse thyroid tissue labeling PAX8 with ab239363 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in mouse thyroid (PMID : 21317881). The section was incubated with ab239363 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thyroid staining of Thyroid Peroxidase/TPO with ab278525 at a 1/5000 (0.109 μg/ml) dilution, PAX8 with ab239363 at 1/2000 (0.254 μg/ml) dilution, and CaSR with ab259846 at 1/5000 (0.103 μg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Thyroid peroxidase (green; Opal™ 520), anti-PAX8 (magenta; Opal™ 690) and anti-CaSR (gray; Opal™ 570) on mouse thyroid.
Panel B : anti-Thyroid peroxidase showed cytoplasmic and membranous staining on mouse thyroid.
Panel C : anti-PAX8 showed nucleus staining on mouse thyroid.
Panel D : anti-CaSR staining parafollicular cells.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab278525, ab239363 and ab259846 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thyroid staining of Thyroid Peroxidase/TPO with ab278525 at a 1/5000 (0.109 μg/ml) dilution, PAX8 with ab239363 at 1/2000 (0.254 μg/ml) dilution, and Collagen I with ab270993 at a 1/500 (1.152 μg/ml) dilution, followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Thyroid peroxidase (green; Opal™ 520), anti-PAX8 (magenta; Opal™ 690) and anti-Collagen I (gray; Opal™ 570) on mouse thyroid.
Panel B : anti-Thyroid peroxidase showed cytoplasmic and membranous staining on mouse thyroid.
Panel C : anti-PAX8 showed nucleus staining on mouse thyroid.
Panel D : anti-Collagen I staining connective tissues in mouse thyroid.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab278525, ab239363 and ab270993 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : The antibody detects isoform of PAX8

The molecular weight observed is consistent with what has been described in the literature (PMID : 21602887)

This blot was developed using a higher sensitivity ECL substrate.

Fresh lysate was used in this Lane 2.

Lane 1 : 127 seconds Lane 2 : 59 seconds

Exposure time :

All lanes:

Western blot - Anti-PAX8 antibody [EPR23508-20] (<a href='/ko/products/primary-antibodies/pax8-antibody-epr23508-20-ab239363'>ab239363</a>) at 1/1000 dilution

Lane 1:

SK-OV-3 (human ovarian cancer epithelial cell), whole cell lysate at 20 µg

Lane 2:

NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 48 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

"This data was developed using the same antibody clone in a different buffer formulation (ab239363). 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 ab239363 [EPR23508-20]. 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 [EPR23508-20] - BSA and Azide free (AB275259)

"This data was developed using the same antibody clone in a different buffer formulation (ab239363). 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 ab239363 [EPR23508-20]. 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 [EPR23508-20] - BSA and Azide free (AB275259)

"This data was developed using the same antibody clone in a different buffer formulation (ab239363). 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 ab239363 [EPR23508-20]. 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."

• WB

Lab

Western blot - Anti-PAX8 antibody [EPR23508-20] - BSA and Azide free (AB275259)

This data was developed using ab239363, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : The molecular weight observed is consistent with what has been described in the literature (PMID : 21602887)

This blot was developed using a higher sensitivity ECL substrate.

3 minutes

Exposure time :

All lanes:

Western blot - Anti-PAX8 antibody [EPR23508-20] (<a href='/ko/products/primary-antibodies/pax8-antibody-epr23508-20-ab239363'>ab239363</a>) at 1/1000 dilution

All lanes:

Mouse E18 kidney tissue lysate at 60 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 48 kDa

false