Recombinant
RabMAb

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

Rabbit recombinant monoclonal Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Human. Cited in 47 publication(s).

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

      

    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:
    • Morohoshi A  et al. The ubiquitin ligase subunit ß-TrCP in Sertoli cells is essential for spermatogenesis in mice. Dev Biol 445:178-188 (2019). Read more (PubMed: 30391586) »
    • Krueger K  et al. Deletion of an intronic HIF-2a binding site suppresses hypoxia-induced WT1 expression. Biochim Biophys Acta Gene Regul Mech 1862:71-83 (2019). Read more (PubMed: 30468780) »
    See all 59 Publications for this product

    Customer reviews and Q&As

    21-30 of 30 Abreviews or Q&A

    Question
    Answer

    The primary and secondary antibody used in the flow cytometry experiment with the Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901) were diluted in PBS with 1% FBS.

    Read More
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (mouse testes)
    Specification
    mouse testes
    Fixative
    4% PFA
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/citrate sodium
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Dr. Qing Wen

    Verified customer

    Submitted Mar 06 2013

    Answer

    Vielen Dank für Ihre Email. Ich habe die Anschrift geändert.

    Ich freue mich Ihnen mitteilen zu können, dass die Konjugation des Antikörpers nun abgeschlossen ist und ab140484 im Moment in der Aufreinigungsphase ist.

    Wir werden Ihnen ab140484 bald zuschicken und möchten uns herzlich für Ihre Geduld bedanken.

    Read More

    Question
    Answer

    Thank you for your reply.

    Your credit note ID is ***.

    I am sorry that this antibody did not perform as expected. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone at 888-772-2226. Please refer to the credit note ID in any correspondence with our accounting department.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Question
    Answer

    Thank you very much for contacting us and letting us know about the trouble with the vial of this antibody.

    Thank you also for sending the details of your protocol and the images. I do have a couple of additional questions, so that I completely understand the situation:

    1) How long do you perform the heat-mediated antigen retrieval, and in what kind of buffer (citrate, EDTA, etc)?

    2) Have you changed any details of your protocol between the old and new vials?

    3) What was the lot number of the vial that worked in the past?

    We do not test this antibody with rat samplse, and it is not covered by our guarantee. However since it has worked for you in the past, I'd be happy to send you a replacement vial. Would you be able to submit an Abreview online, and we may be able to add rat to the list of our guaranteed species?

    I look forward to hearing from you. Please let me know if you would like to receive a replacment vial and I will send this to you promptly.

    Read More

    Answer

    Thank you very much for contacting us and letting us know about the trouble with the vial of this antibody.

    Thank you also for sending the details of your protocol and the images. I do have a couple of additional questions, so that I completely understand the situation:

    1) How long do you perform the heat-mediated antigen retrieval, and in what kind of buffer (citrate, EDTA, etc)?

    2) Have you changed any details of your protocol between the old and new vials?

    3) What was the lot number of the vial that worked in the past?

    We do not test this antibody with rat samplse, and it is not covered by our guarantee. However since it has worked for you in the past, I'd be happy to send you a replacement vial. Would you be able to submit an Abreview online, and we may be able to add rat to the list of our guaranteed species?

    I look forward to hearing from you. Please let me know if you would like to receive a replacment vial and I will send this to you promptly.

    Read More

    Answer

    Thank you for your inquiry and your patience.

    I have now confirmation that we will be able to produce ab89901 as a directly FITC conjugated version.

    I have to confirm that this will take 8- 11 weeks to be produced. I apologize for the long lead time.



    We are thinking about including this antibody version in our catalog andin this casefuture orders will be easier and faster.

    I hope this information is helpful. Please let me know if you are interested in purchasing this antibody.

    Read More
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (heart (epicardial expression))
    Specification
    heart (epicardial expression)
    Fixative
    Formaldehyde
    Permeabilization
    Yes - 0.5% Tx100 in PBS
    Blocking step
    1% goat serum 10% BSA 0.1% Tx100 PBS as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Jun 18 2012

    Answer

    Thank you for contacting Abcam regarding ab89901

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Kidney)
    Specification
    Kidney
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: sodium citrate
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C

    Dr. R Berahovich

    Verified customer

    Submitted Nov 23 2011

    21-30 of 30 Abreviews or Q&A

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