Recombinant
RabMAb

Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)

Overview

  • Product name
    Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2]
    See all Wilms Tumor Protein primary antibodies
  • Description
    Rabbit monoclonal [CAN-R9(IHC)-56-2] to Wilms Tumor Protein
  • Host species
    Rabbit
  • Specificity
    Expression levels of the target protein vary with sample type and some optimisation may be required.
  • Tested applications
    Suitable for: IHC-Fr, Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment (GST-tag) within Human Wilms Tumor Protein aa 50-250. The exact sequence is proprietary.
    Database link: P19544
    (Peptide available as ab208102)

  • Positive control
    • WB: Human testis whole cell lysate. K562 and Ramos cell lysate. IHC-P: Human kidney and Wilms tumor tissues. Human ovarian serous adenocarcinoma tissue. ICC/IF: K562 cells. Flow Cyt: K562 cells. IHC-Fr: Human embryonic kidney tissue.
  • General notes

     

     

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab89901 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr 1/250.
Flow Cyt 1/50.

For unpurified use at 1/3.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/500 - 1/1000. Predicted molecular weight: 55 kDa.

For unpurified use at 1/100.

IHC-P 1/300. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/30.

ICC/IF 1/50.

For unpurified use at 1/5.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Transcription factor that plays an important role in cellular development and cell survival. Regulates the expression of numerous target genes, including EPO. Plays an essential role for development of the urogenital system. Recognizes and binds to the DNA sequence 5'-CGCCCCCGC-3'. 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. Isoforms containing the KTS motif may bind mRNA and play a role in mRNA metabolism or splicing. Isoform 1 has lower affinity for DNA, and can bind RNA.
    • Tissue specificity
      Expressed in the kidney and a subset of hematopoietic cells.
    • Involvement in disease
      Defects in WT1 are the cause of Frasier syndrome (FS) [MIM:136680]. FS is characterized by a slowly progressing nephropathy leading to renal failure in adolescence or early adulthood, male pseudohermaphroditism, and no Wilms tumor. As for histological findings of the kidneys, focal glomerular sclerosis is often observed. There is phenotypic overlap with Denys-Drash syndrome. Inheritance is autosomal dominant.
      Defects in WT1 are the cause of Wilms tumor 1 (WT1) [MIM:194070]. WT is an embryonal malignancy of the kidney that affects approximately 1 in 10'000 infants and young children. It occurs both in sporadic and hereditary forms.
      Defects in WT1 are the cause of Denys-Drash syndrome (DDS) [MIM:194080]. DDS is a typical nephropathy characterized by diffuse mesangial sclerosis, genital abnormalities, and/or Wilms tumor. There is phenotypic overlap with WAGR syndrome and Frasier syndrome. Inheritance is autosomal dominant, but most cases are sporadic.
      Defects in WT1 are the cause of nephrotic syndrome type 4 (NPHS4) [MIM:256370]. A renal disease characterized clinically by proteinuria, hypoalbuminemia, hyperlipidemia and edema. Kidney biopsies show non-specific histologic changes such as focal segmental glomerulosclerosis and diffuse mesangial proliferation. Some affected individuals have an inherited steroid-resistant form and progress to end-stage renal failure. Most patients with NPHS4 show diffuse mesangial sclerosis on renal biopsy, which is a pathologic entity characterized by mesangial matrix expansion with no mesangial hypercellularity, hypertrophy of the podocytes, vacuolized podocytes, thickened basement membranes, and diminished patency of the capillary lumen.
      Defects in WT1 are a cause of Meacham syndrome (MEACHS) [MIM:608978]. Meacham syndrome is a rare sporadically occurring multiple malformation syndrome characterized by male pseudohermaphroditism with abnormal internal female genitalia comprising a uterus and double or septate vagina, complex congenital heart defect and diaphragmatic abnormalities.
      Note=A chromosomal aberration involving WT1 may be a cause of desmoplastic small round cell tumor (DSRCT). Translocation t(11;22)(p13;q12) with EWSR1.
    • Sequence similarities
      Belongs to the EGR C2H2-type zinc-finger protein family.
      Contains 4 C2H2-type zinc fingers.
    • Cellular localization
      Nucleus. Cytoplasm. Shuttles between nucleus and cytoplasm; Nucleus > nucleoplasm and Nucleus speckle.
    • Information by UniProt
    • Database links
    • Alternative names
      • WIT 2 antibody
      • WT 1 antibody
      • AWT1 antibody
      • FWT1 antibody
      • GUD antibody
      • NPHS4 antibody
      • WAGR antibody
      • Wilms tumor 1 antibody
      • Wilms Tumor antibody
      • Wilms tumor protein antibody
      • Wilms' tumor gene antibody
      • Wilms' tumor protein antibody
      • WIT2 antibody
      • WT antibody
      • WT1 antibody
      • WT1_HUMAN antibody
      • WT33 antibody
      see all

    Images

    • 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.

    • 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.

    • 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.

    • Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/100 dilution (unpurified) + K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 55 kDa
      Observed band size: 55 kDa



      Blocking buffer and concentration: 5% NFDM/TBST.

    • Immunocytochemsitry/Immunofluorescence analysis of K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells lablling Wilms Tumor Protein (green) with unpurified 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).

    • ab89901 staining Wilms Tumor Protein in human embryonic kidney tissue sections by Immunohistochemistry (IHC-Fr - frozen sections).

      Tissue was fixed with formaldehyde, permeabilized with 0.25% Triton X and blocked with 3% BSA and donkey antisera for 60 minutes at 23°C. Samples were incubated with primary antibody (1/1000 in PBS + 0.1% Triton + 0.1% BSA) for 16 hours at 4°C. A Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

      See Abreview

    • 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 4ºC. The tissues were blocked with 10% serum for 30 minutes at 25ºC. The secondary used was a Donkey CY3® conjugate at a 1/200 dilution.

      See Abreview

    • 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.

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

      Predicted band size: 55 kDa
      Observed band size: 55 kDa



      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

       

       

    • 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 Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) / Black was used as the isotype control. Cells without incubation with primary antibody and secondary antibody / Blue was used as the unlabeled control.

    • 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Wilms tumor tissue labeling Wilms Tumor Protein with unpurified ab89901 at 1/250.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal tissue labeling Wilms Tumor Protein with unpurified ab89901 at 1/250.

    • 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).

    • All lanes : 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
      Lane 2 : THP-1 (Human monocytic leukemia monocyte). Whole cell lysates
      Lane 3 : SH-SY5Y (Human neuroblastoma epithelial cell) Whole cell lysates
      Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 55 kDa


      Exposure time: 3 minutes


      WT1 expression varies in different cell lines. 

      Blocking and diluting buffer: 5% NFDM/TBST.

    • Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution (purified) + K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anto-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 55 kDa
      Observed band size: 55 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution (unpurified) + Ramos (Human Burkitt's lymphoma cell line) cell lysate

      Predicted band size: 55 kDa

    References

    This product has been referenced in:
    • O'Brien LL  et al. Transcriptional regulatory control of mammalian nephron progenitors revealed by multi-factor cistromic analysis and genetic studies. PLoS Genet 14:e1007181 (2018). Read more (PubMed: 29377931) »
    • Sereti KI  et al. Analysis of cardiomyocyte clonal expansion during mouse heart development and injury. Nat Commun 9:754 (2018). Read more (PubMed: 29467410) »
    See all 44 Publications for this product

    Customer reviews and Q&As

    1-10 of 29 Abreviews or Q&A

    Answer

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    Read More
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Pig Tissue sections (Heart)
    Permeabilization
    Yes - PBS + Triton 0.1%
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Davide Maselli

    Verified customer

    Submitted Aug 23 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton X-100
    Specification
    HeLa
    Fixative
    Paraformaldehyde

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Aug 10 2018

    Application
    ChIP
    Sample
    Human Cell lysate - nuclear (K562 Human Cell Line)
    Negative control
    PU1 Enhancer
    Specification
    K562 Human Cell Line
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% Formaldehyde
    Positive control
    HMBS, JUN Promoter, NAB2, WNT11A

    Abcam user community

    Verified customer

    Submitted Jun 26 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Whole brain tissue lysate)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (3-8% Bis-Tris)
    Loading amount
    20 µg
    Specification
    Whole brain tissue lysate
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Mar 29 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human pluripotent SC derived epicardial like cells)
    Permeabilization
    Yes - Saponin
    Specification
    Human pluripotent SC derived epicardial like cells
    Blocking step
    Serum as blocking agent for 15 minute(s) · Concentration: 4% · Temperature: 4°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Nov 16 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Breast tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TE pH8
    Permeabilization
    No
    Specification
    Breast tumor
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    10% normal buffered formalin

    Abcam user community

    Verified customer

    Submitted Aug 26 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TE pH 8.0
    Permeabilization
    No
    Specification
    kidney
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Aug 09 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer
    Permeabilization
    Yes - 0.03% Triton for 10min
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 28 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Human foetal heart)
    Permeabilization
    Yes - 0.5% Triton-X
    Specification
    Human foetal heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 07 2016

    1-10 of 29 Abreviews or Q&A

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