Anti-PAX8 antibody [EPR23508-20]

15 product images

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  Immunoprecipitation - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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)(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: abab239363 IP in SK-OV-3 whole cell lysate

  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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] (ab239363)

  Predicted band size: 48 kDa

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  Western blot - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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.

  Exposure time: 3 minutes

  All lanes: Western blot - Anti-PAX8 antibody [EPR23508-20] (ab239363) 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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 48 kDa

• 

  Western blot - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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.

  Exposure time: Lane 1: 127 seconds Lane 2: 59 seconds

  All lanes: Western blot - Anti-PAX8 antibody [EPR23508-20] (ab239363) 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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 48 kDa

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  Immunohistochemistry (Frozen sections) - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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).

• 

  Flow Cytometry (Intracellular) - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 (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 (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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 (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 (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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  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 (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 (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.

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  ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR23508-20] (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 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods."

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  Multiplex immunohistochemistry - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thyroid staining of Thyroid Peroxidase/TPO with Anti-Thyroid Peroxidase/TPO antibody [EPR23574-405] ab278525 at a 1/5000 (0.109 μg/ml) dilution, PAX8 with ab239363 at 1/2000 (0.254 μg/ml) dilution, and CaSR with Anti-CaSR antibody [EPR24050-59] 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 Anti-Thyroid Peroxidase/TPO antibody [EPR23574-405] ab278525, ab239363 and Anti-CaSR antibody [EPR24050-59] 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.

• 

  ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR23508-20] (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 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods."

• 

  ChIC/CUT&RUN sequencing - Anti-PAX8 antibody [EPR23508-20] (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 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods."

• 

  Multiplex immunohistochemistry - Anti-PAX8 antibody [EPR23508-20] (ab239363)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thyroid staining of Thyroid Peroxidase/TPO with Anti-Thyroid Peroxidase/TPO antibody [EPR23574-405] 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 Anti-Collagen I antibody [EPR24331-53] 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 Anti-Thyroid Peroxidase/TPO antibody [EPR23574-405] ab278525, ab239363 and Anti-Collagen I antibody [EPR24331-53] 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.