Recombinant Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] (ab89901)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CAN-R9(IHC)-56-2] to Wilms Tumor Protein
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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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: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
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Positive control
- WB: Human testis whole cell lysate. K562, Ramos, THP-1 and HeLa whole cell lysates. IHC-P: Human kidney, Wilms tumor, ovarian serous adenocarcinoma and fetal tissues; Rat and mouse testis tissues. ICC/IF: K562 cells. Flow Cyt (intra): K562 cells. IHC-Fr: Human embryonic kidney tissue.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
CAN-R9(IHC)-56-2 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- 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)
- Anti-Wilms Tumor Protein antibody [CAN-R9(IHC)-56-2] - BSA and Azide free (ab216646)
- 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)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
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 |
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WB | (4) |
1/500 - 1/1000. Predicted molecular weight: 55 kDa.
For unpurified use at 1/100. |
IHC-P | (14) |
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. |
Flow Cyt (Intra) |
1/50.
For unpurified use at 1/3. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF | (6) |
1/50.
For unpurified use at 1/5. |
Notes |
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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. |
Flow Cyt (Intra)
1/50. For unpurified use at 1/3. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/50. For unpurified use at 1/5. |
Target
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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
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Database links
- Entrez Gene: 7490 Human
- Entrez Gene: 22431 Mouse
- Omim: 607102 Human
- SwissProt: P19544 Human
- SwissProt: P22561 Mouse
- Unigene: 591980 Human
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Alternative names
- WIT 2 antibody
- WT 1 antibody
- AWT1 antibody
see all
Images
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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.
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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).
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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 kDaExposure time: Left image: 3 minutes
Right image: 15 secondsThis antibody shows low affinity in mouse sample.
Blocking and diluting buffer: 5% NFDM/TBST
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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, 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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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 kDaBlocking buffer and concentration: 5% NFDM/TBST.
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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 minutesWT1 expression varies in different cell lines.
Blocking and diluting buffer: 5% NFDM/TBST.
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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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (208)
ab89901 has been referenced in 208 publications.
- Matoba K et al. ROCK2-induced metabolic rewiring in diabetic podocytopathy. Commun Biol 5:341 (2022). PubMed: 35396346
- Floy ME et al. Directed differentiation of human pluripotent stem cells to epicardial-derived fibroblasts. STAR Protoc 3:101275 (2022). PubMed: 35403005
- Jablonowski CM et al. TERT Expression in Wilms Tumor Is Regulated by Promoter Mutation or Hypermethylation, WT1, and N-MYC. Cancers (Basel) 14:N/A (2022). PubMed: 35406427
- Nystrom SE et al. JAK inhibitor blocks COVID-19 cytokine-induced JAK/STAT/APOL1 signaling in glomerular cells and podocytopathy in human kidney organoids. JCI Insight 7:N/A (2022). PubMed: 35472001
- Malolina EA et al. Establishment of a pure culture of immature Sertoli cells by PDGFRA staining and cell sorting. Mol Reprod Dev 89:243-255 (2022). PubMed: 35478364