Anti-Wilms Tumor Protein antibody [EPR23963-116]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized OVCAR-3 (human ovary adenocarcinoma epithelial cell) cells labelling Wilms tumor protein with ab267377 at 1/50 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 OVCAR-3 cells. Negative control : LnCap (PMID : 11299720). 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded Human testis tissue sections labeling DKKL1 with ab316308 at a 1/2000 dilutions (B), Wilms Tumor Protein with ab275970 at a 1/1200 dilutions (C), and SAGE1 with ab233388 at a 1/250 dilutions (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-DKKL1 (red; Opal™ 570), anti-Wilms Tumor Protein (green; Opal™ 520) and anti-SAGE1 (gray; Opal™ 690) on human testis.
Panel B : anti-DKKL1 staining mature sperm cells in human testis.
Panel C : anti-Wilms Tumor Protein staining sertoli cells in human testis.
Panel D : anti-SAGE1 staining spermatogonia in human testis.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The section was incubated in three rounds of staining : in the order of ab316308, ab275970, and ab233388 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

This data was developed using ab275970, the same antibody clone in a different buffer formulation.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on Sertoli cells of human testis (PMID : 17229929). The section was incubated with ab267377 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 Goat Anti-Rabbit IgG H&L (HRP).

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-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on human ovarian carcinoma (PMID : 16547468). The section was incubated with ab267377 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 Goat Anti-Rabbit IgG H&L (HRP).

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-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on human renal glomeruli (PMID : 27922671). The section was incubated with ab267377 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 LeicaDS9800 (Bond^® Polymer Refine Detection).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections Rat testis tissue labelling Lin28A with ab279647 at 1 : 1000 dilution (B), Wilms Tumor Protein with ab267377 at 1 : 500 dilution (C) and DKKL1 with ab316308 at 1 : 2000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Lin28A (green; Opal™520), anti-Wilms Tumor Protein (magenta; Opal™690) and anti-DKKL1 (gray; Opal™570) on rat testis.

Panel B : anti-Lin28A staining undifferentiated spermatogonia in rat testis.

Panel C : ant-Wilms Tumor Protein staining Sertoli cells in rat testis.

Panel D : ant-DKKL11 staining mature sperm cells in rat testis.

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

The section was incubated in three rounds of staining : in the order of ab279647, ab267377 and ab316308 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™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on Sertoli cells of rat testis (PMID : 17229929). The section was incubated with ab267377 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 LeicaDS9800 (Bond^® Polymer Refine Detection) was used.

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-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on Sertoli cells of mouse testis (PMID : 17229929). The section was incubated with ab267377 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 LeicaDS9800 (Bond^® Polymer Refine Detection).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse testis tissue staining Laminin alpha 1 with ab307542 at a 1/100 dilution, ab267377 anti-Wilms tumor protein used at 1/2000 dilution and ab323317 anti-Oligodendrocyte Specific Protein used at a 1/500 dilution.

Panel A : anti-Laminin alpha 1 (green; Opal™520), anti-Wilms tumor protein (magenta; Opal™690) and anti Oligodendrocyte Specific Protein (gray; Opal™570) on mouse testis.

Panel B : anti-Laminin alpha 1 staining basement membrane in mouse testis.

Panel C : anti-Wilms tumor protein staining Sertoli cells in mouse testis.

Panel D : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in mouse testis.

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

The section was incubated in three rounds of staining : in the order of ab307542, ab267377 and ab323317 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™ 4-color kit. 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat testis tissue staining Laminin alpha 1 with ab307542 at a 1/100 dilution, ab267377 anti-Wilms tumor protein used at 1/2000 dilution and ab323317 anti-Oligodendrocyte Specific Protein used at a 1/500 dilution.

Panel A : anti-Laminin alpha 1 (green; Opal™520), anti-Wilms tumor protein (magenta; Opal™690) and anti Oligodendrocyte Specific Protein (gray; Opal™570) on rat testis.

Panel B : anti-Laminin alpha 1 staining basement membrane in rat testis.

Panel C : anti-Wilms tumor protein staining Sertoli cells in rat testis.

Panel D : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in rat testis.

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

The section was incubated in three rounds of staining : in the order of ab307542, ab267377 and ab323317 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™ 4-color kit. 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections Mouse testis tissue labelling Lin28A with ab279647 at 1 : 1000 dilution (B), Wilms Tumor Protein with ab267377 at 1 : 500 dilution (C) and DKKL1 with ab316308 at 1 : 2000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Lin28A (green; Opal™520), anti-Wilms Tumor Protein (magenta; Opal™690) and anti-DKKL1 (gray; Opal™570) on rat testis.

Panel B : anti-Lin28A staining undifferentiated spermatogonia in rat testis.

Panel C : ant-Wilms Tumor Protein staining Sertoli cells in rat testis.

Panel D : ant-DKKL11 staining mature sperm cells in rat testis.

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

The section was incubated in three rounds of staining : in the order of ab279647, ab267377 and ab316308 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™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

Lanes 1 - 4 : Merged signal (red and green). Green - ab267377 observed at 50-60 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab267377 was shown to react with Wilms Tumor Protein in wild-type HAP1 cells in Western blot with loss of signal observed in Wt1 knockout sample. Wild-type HAP1 and Wt1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab267377 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

Wt1 knockout HAP1 cell lysate at 20 µg

Lane 3:

K-562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg

Predicted band size: 49 kDa

Observed band size: 50-60 kDa

false

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 10438524).

Negative control : LNCaP (PMID : 11299720).

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 49 kDa

Observed band size: 52 kDa,54 kDa

false

• WB

Lab

Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (AB267377)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 10438524).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 49 kDa

Observed band size: 52 kDa,54 kDa

false