Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)

Rabbit Recombinant Monoclonal Wilms Tumor Protein antibody. Suitable for ICC/IF, mIHC, WB, IHC-P and reacts with Human, Mouse, Rat samples. Cited in 3 publications.

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Images

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

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	mIHC	IP	Flow Cyt	WB	IHC-Fr	IHC-P
Human	Tested	Tested	Not recommended	Not recommended	Tested	Not recommended	Tested
Mouse	Expected	Expected	Not recommended	Not recommended	Tested	Not recommended	Tested
Rat	Expected	Expected	Not recommended	Not recommended	Tested	Not recommended	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information WT1

Function

Transcription factor that plays an important role in cellular development and cell survival (PubMed:7862533). Recognizes and binds to the DNA sequence 5'-GCG(T/G)GGGCG-3' (PubMed:17716689, PubMed:25258363, PubMed:7862533). Regulates the expression of numerous target genes, including EPO. Plays an essential role for development of the urogenital system. It has a tumor suppressor as well as an oncogenic role in tumor formation. Function may be isoform-specific: isoforms lacking the KTS motif may act as transcription factors (PubMed:15520190). Isoforms containing the KTS motif may bind mRNA and play a role in mRNA metabolism or splicing (PubMed:16934801). Isoform 1 has lower affinity for DNA, and can bind RNA (PubMed:19123921).

Alternative names

Wilms tumor protein, WT33, WT1

Recommended products

Rabbit Recombinant Monoclonal Wilms Tumor Protein antibody. Suitable for ICC/IF, mIHC, WB, IHC-P and reacts with Human, Mouse, Rat samples. Cited in 3 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23963-116
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Wilms Tumor Protein commonly known as WT1 is a critical transcription factor implicated in several cellular processes. It is also identified by other names such as Wilms Tumor Suppressor and WT33. WT1 protein has a mass of approximately 52-54 kDa. It is broadly expressed in various tissues with high levels in the developing kidney gonads and certain mesothelial tissues. Its expression plays an important role in organ development and cellular differentiation.

Biological function summary

The WT1 protein regulates gene expression by binding to specific DNA sequences influencing cell growth and differentiation. WT1 acts as a part of larger protein complexes interacting with other transcription factors and co-regulators. It plays essential roles in the development of the urogenital system and in the maintenance of mesothelial cells. The WT1 protein is essential for normal kidney and gonadal development highlighting its significance in embryogenesis.

Pathways

WT1 integration involves multiple cellular signaling cascades. It is notably engaged in the Wnt and PI3K/AKT signaling pathways which are critical for cell proliferation and survival. In these pathways WT1 interaction with proteins such as β-catenin and PI3K subunits modulates cell fate decisions. This involvement in signaling networks exemplifies its importance in cellular homeostasis and response to environmental cues.

Associated diseases and disorders

WT1 mutations or dysregulation associate strongly with specific pathologies particularly Wilms tumor and acute myeloid leukemia (AML). In Wilms tumor WT1 acts as a tumor suppressor gene and its loss of function leads to tumorigenesis in the kidney. In AML aberrant WT1 expression affects normal hematopoiesis and is often linked to poor prognosis. Interaction with proteins like BSA in immunohistochemistry diagnostics highlights WT1’s significance as a biomarker for these diseases.

Product promise

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10 product images

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 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 OVCAR-3 cells. Negative control: LnCap (PMID:11299720). 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.
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.
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 Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 50-60 kDa
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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 49 kDa
Observed band size: 52 kDa, 54 kDa
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.
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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 49 kDa
Observed band size: 52 kDa, 54 kDa
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.
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.
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.
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 Anti-DKKL1 antibody [EPR25721-68] ab316308 at a 1/2000 dilutions (B), Wilms Tumor Protein with Anti-Wilms Tumor Protein antibody [EPR23963-116] - BSA and Azide free ab275970 at a 1/1200 dilutions (C), and SAGE1 with Anti-SAGE1 antibody [EPR21747] 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 Anti-DKKL1 antibody [EPR25721-68] ab316308, Anti-Wilms Tumor Protein antibody [EPR23963-116] - BSA and Azide free ab275970, and Anti-SAGE1 antibody [EPR21747] 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 Anti-Wilms Tumor Protein antibody [EPR23963-116] - BSA and Azide free ab275970, the same antibody clone in a different buffer formulation.