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

Overview

  • Product nameAnti-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
  • Tested applicationsSuitable for: IHC-Fr, Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) (GST-tag) corresponding to Human Wilms Tumor Protein aa 50-250.
    Database link: P19544

  • Positive control
    • WB: K562 and Ramos cell lysate. IHC-P: Human kidney and Wilms tumor tissues. ICC/IF: K562 cells. Flow Cyt: K562 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:

    Anti-Wilms Tumor Protein antibody (Alexa Fluor® 647) [CAN-R9(IHC)-56-2] (ab202639)

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 1/1000. Predicted molecular weight: 55 kDa.

For unpurified use at 1/100.

IHC-P 1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/30.

ICC/IF 1/50.

For unpurified use at 1/5.

  • Application notesIs unsuitable for IP.
  • Target

    • FunctionTranscription 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 specificityExpressed in the kidney and a subset of hematopoietic cells.
    • Involvement in diseaseDefects 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 similaritiesBelongs to the EGR C2H2-type zinc-finger protein family.
      Contains 4 C2H2-type zinc fingers.
    • Cellular localizationNucleus. 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

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

    • Immunohistochemical analysis of fromaldehyde fixed, paraffin embedded rat testis tissue sections, labelling 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 Human 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

    • Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/100 dilution (unpurified) + K562 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.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) at 1/1000 dilution (purified) + K562 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 buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

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

      Predicted band size : 55 kDa
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Wilms Tumor Protein with unpurified ab89901 at 1/30. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Wilms Tumor Protein with purified ab89901 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

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

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

    • Immunocytochemsitry/Immunofluorescence analysis of K562 cells labelling 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).

    • Immunocytochemsitry/Immunofluorescence analysis of K562 cells labelling 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).

    • Overlay histogram showing K562 cells stained with unpurified ab89901 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab89901, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Flow cytometry analysis of K562 cells labelling Wilms Tumor Protein with unpurified ab89901 at 1/3 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

    • Flow cytometry analysis of K562 cells labelling Wilms Tumor Protein with purified ab89901 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

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

    This product has been referenced in:
    • Liu M  et al. Recovery of fertility in quinestrol- or diethylstilbestrol (DES)-treated mice: implications for rodent management. Integr Zool N/A:N/A (2016). Read more (PubMed: 27611741) »
    • Risebro CA  et al. Characterisation of the human embryonic and foetal epicardium during heart development. Development 142:3630-6 (2015). IHC ; Human . Read more (PubMed: 26395486) »

    See all 12 Publications for this product

    Product Wall

    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
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    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
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    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
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    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
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    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
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    Submitted Jul 07 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 (testis (Sertoli cells))
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA pH9
    Permeabilization Yes - Tween-20
    Specification testis (Sertoli cells)
    Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative Paraformaldehyde
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    Submitted May 19 2016

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

    Submitted May 12 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Sample Human Tissue sections (Human embryonic kidney)
    Permeabilization Yes - 0.25% Triton X
    Specification Human embryonic kidney
    Blocking step BSA and donkey antisera as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
    Fixative Formaldehyde
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    Submitted Apr 04 2016

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Rat Tissue sections (Testis)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM Sodium Citrate 0,05% Tween 20
    Permeabilization No
    Specification Testis
    Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative Formaldehyde
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    Verified customer

    Submitted Oct 19 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.01M citric acide/sodium citrate
    Sample Sheep Tissue sections (1-2 month old testis)
    Specification 1-2 month old testis
    Permeabilization No
    Fixative 4%PFA
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    Submitted Jan 14 2015

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