Anti-p63 antibody [EPR5701]

7 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p63 antibody [EPR5701] (ab124762)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling p63 with purified ab124762 at 1/5000 dilution (0.16 μg/mL). Heat mediated antigen retrieval using Bond™™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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  Western blot - Anti-p63 antibody [EPR5701] (ab124762)

  Blocking/Diluting buffer: 5% NFDM/TBST
  

  The bands observed are consistent with what have been described in PMID 30649915 as isoforms of p63.

  All lanes: Western blot - Anti-p63 antibody [EPR5701] (ab124762) at 1/1000 dilution

  Lane 1: HaCaT (Human skin keratinocyte) whole cell lysates at 20 µg

  Lane 2: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 3: Mouse skin lysates at 20 µg

  Lane 4: Mouse thymus lysates at 20 µg

  Lane 5: Mouse bladder lysates at 20 µg

  Lane 6: Rat skin lysates at 20 µg

  Lane 7: Rat bladder lysates at 20 µg

  Lane 8: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 77 kDa

  Observed band size: 37-75 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-p63 antibody [EPR5701] (ab124762)

  Immunocytochemistry/ Immunofluorescence analysis of A431(Human epidermoid carcinoma epithelial cell) cells labeling p63 with Purified ab124762 at 1/200 dilution (4 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  Flow Cytometry (Intracellular) - Anti-p63 antibody [EPR5701] (ab124762)

  Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling p63 with Purified ab124762 at 1/80 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

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  This image is courtesy of a customer review submitted by Manuel Chacon

  Immunocytochemistry/ Immunofluorescence - Anti-p63 antibody [EPR5701] (ab124762)

  Unpurified ab124762 staining p63 in Human corneal limbal epithelial cells (primary culture) by ICC/IF (immunocytochemistry/immunofluorescence). Cells were fixed with methanol and permeabilized with 0.3% Triton X-100 for 5 minutes. Samples were incubated with primary antibody (1/100 in PBS + 10% Goat serum) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

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  Immunohistochemistry - Anti-p63 antibody [EPR5701] (ab124762)

  Immunohistochemical analysis of formalin fixed paraffin embedded human prostate labelling p63 with ab124762 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

  ab124762 anti-p63 [EPR5701] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

  Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

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  Multiplex immunohistochemistry - Anti-p63 antibody [EPR5701] (ab124762)

  Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-p63 (ab124762, magenta; Opal™690), anti-Cytokeratin 5 (Anti-Cytokeratin 5 antibody [SP27] - BSA and Azide free ab236216, green; Opal™520) and anti-Prostate Specific Antigen (Anti-Prostate Specific Antigen antibody [EP1588Y] ab76113, red; Opal™570) on human prostate gland tissue. Panel B: anti-Prostate Specific Antigen stained on luminal cells. Panel C: anti-Cytokeratin 5 stained on cytoplasm of basal cells. Panel D: anti-p63 stained on nucleus of basal cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab124762 (1/5000), Anti-Cytokeratin 5 antibody [SP27] - BSA and Azide free ab236216 (1/400), and Anti-Prostate Specific Antigen antibody [EP1588Y] ab76113 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. 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 solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.