Name: Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2]
SKU: ab89901
Rating: 5 (42 reviews)

Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] ab89901 is a rabbit monoclonal antibody that is used in Wilms Tumor Protein western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone CAN-R9(IHC)-56-2 is the most widely used clone for Wilms Tumor Protein on the market
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your Wilms Tumor Protein staining

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (AB89901), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Not recommended	Tested	Tested	Tested
Mouse	Expected	Not recommended	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/300	Notes For unpurified use at 1/30. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

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

Not recommended
Not recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500 - 1/1000	Notes For unpurified use at 1/100.
Species Human	Dilution info 1/500 - 1/1000	Notes For unpurified use at 1/100.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes For unpurified use at 1/5.

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes For unpurified use at 1/3. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

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

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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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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: CAN-R9(IHC)-56-2
Purification technique: Affinity purification Protein A
Specificity: Expression levels of the target protein vary with sample type and some optimisation may be required.
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

Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) was first used in a scientific publication in 2011 and has been cited over 252 times in peer reviewed journals. It's performance in IHC in mouse and human samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) has 40 independent reviews from customers.

Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) specifically detects Wilms Tumor Protein (UniProt ID: P19544; Molecular weight: 50kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone CAN-R9(IHC)-56-2 - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free ab216646.

Antibody clone CAN-R9(IHC)-56-2 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 488, Alexa Fluor^® 647, FITC, Alexa Fluor^® 555 (Alexa Fluor® 488 Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] ab202635, Alexa Fluor® 647 Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] ab202639, FITC Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] ab223934, Alexa Fluor® 555 Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] ab314587).

Wilms Tumor Protein (WT1) is a tumor-associated antigen that is overexpressed in various cancers, making it a target for immunotherapeutic approaches. It plays a role in tumor progression and immune evasion, influencing the tumor microenvironment and the effectiveness of immune responses.

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

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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15 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
ab89901 staining Wilms Tumor Protein in paraffin-embedded mouse testis tissue sections by Immunohistochemistry.
Antigen retrieval was by heat mediation using citrate buffer (pH 6.0). Samples were incubated with primary antibody at 1:500 dilution (0.49 μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Nuclear staining on Sertoli cells in mouse testis (PMID: 21863216) is observed.
Adams et al PLoS Genet. 2018 Feb 20;14(2):e1007233. doi: 10.1371/journal.pgen.1007233. eCollection 2018 Feb. Fig S1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Spermatogenic phenotypes of Rnf8;Scml2-dKO mice.
Immunostaining of testicular sections with WT1 (ab89901), a marker of Sertoli cells, and PLZF, a marker of undifferentiated spermatogonia.
For preparation of testicular paraffin blocks, testes of mutants and littermate controls were fixed with 4% paraformaldehyde at 4°C overnight. Testes were then dehydrated and embedded in paraffin. For histological analyses, 6 μm-thick paraffin sections were deparaffinized and autoclaved in Target Retrieval Solution at 121°C for 20 min. The sections were blocked with for 10 min at room temperature, and then incubated with primary antibodies at 4°C overnight.
Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Exposure time: Left image: 3 minutes
Right image: 15 seconds
This antibody shows low affinity in mouse sample.
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/500 dilution
Lane 1: Human testis whole cell lysate. at 15 µg
Lane 2: K562 (Human T cell leukemia T lymphocyte) whole cell lysates at 4 µg
Lane 3: Mouse testis whole cell lysates at 15 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: 49 kDa
Observed band size: 55 kDa
Flow Cytometry (Intracellular) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Intracellular Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Wilms Tumor Protein with ab89901 at 1/200 dilution (0.1 μg)/ Red.
Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black was used as the isotype control. Cells without incubation with primary antibody and secondary antibody / Blue was used as the unlabeled control.
Verma et al PLoS One. 2018 Jun 20;13(6):e0198013. doi: 10.1371/journal.pone.0198013. eCollection 2018. Fig S1. Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Podocyte counting using WT1 staining.
Mouse kidney sections were stained with WT1 antibody ab89901 using immunohistochemistry. (A) Each glomeruli was identified and numbered using ImagePro premier imaging software. (B) A tracing was applied along the Bowman's capsule to assess the surface area of the glomerulus. (C) Each WT1 positive nucleus was identified and the nuclear size assessed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
ab89901 staining Wilms Tumor Protein in paraffin-embedded human ovarian serous adenocarcinoma tissue sections by Immunohistochemistry.
Antigen retrieval was by heat mediation using citrate buffer (pH 6.0) was performed. Samples were incubated with primary antibody at 1:500 dilution (0.49 μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Nuclear staining on human ovarian serous adenocarcinoma (PMID: 11939727) is observed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
ab89901 staining Wilms Tumor Protein in paraffin-embedded human kidney tissue sections by Immunohistochemistry.
Antigen retrieval was by heat mediation using citrate buffer (pH 6.0). Samples were incubated with primary antibody at 1:500 dilution (0.49 μg/ml). A ready to use Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Nuclear staining on human kidney glomerulus (PMID: 12898605) is observed.
This image is courtesy of an anonymous customer review.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Immunohistochemical analysis of fromaldehyde fixed, paraffin embedded rat testis tissue sections, labeling Wilms Tumor Protein using ab89901.
Heat mediated antigen retrival was performed using 10 mM sodium citrate and 0.05% Tween-20. Tissue sections were incubated with ab89901 at a 1/50 dilution for 12 hours at 4°C. The tissues were blocked with 10% serum for 30 minutes at 25°C. The secondary used was a Donkey CY3^® conjugate at a 1/200 dilution.
Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Blocking buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/100 dilution
All lanes: K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 10 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 55 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Immunocytochemsitry/Immunofluorescence analysis of K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling Wilms Tumor Protein (green) with purified ab89901 at 1/50.
Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).
Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
WT1 expression varies in different cell lines.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/5000 dilution
Lane 1: K-562 (Human chronic myelogenous leukemia lymphoblast). Whole cell lysates at 20 µg
Lane 2: THP-1 (Human monocytic leukemia monocyte). Whole cell lysates at 20 µg
Lane 3: SH-SY5Y (Human neuroblastoma epithelial cell) Whole cell lysates at 20 µg
Lane 4: HeLa (Human cervix adenocarcinoma epithelial cell) 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: 49 kDa
Exposure time: 3min
Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution
All lanes: K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 10 µg
Secondary
All lanes: Peroxidase-conjugated goat anto-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 55 kDa
Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
All lanes: Western blot - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution
All lanes: Ramos (Human Burkitt's lymphoma cell line) cell lysate
Predicted band size: 49 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal tissue labeling Wilms Tumor Protein with unpurified ab89901 at 1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Wilms tumor tissue labeling Wilms Tumor Protein with unpurified ab89901 at 1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.