Recombinant Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)
![Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-6-anti-wilms-tumor-protein-antibody-epr23963116-immunocytochemistry-ovcar3-lncap.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23963-116] to Wilms Tumor Protein
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Wilms Tumor Protein antibody [EPR23963-116]
See all Wilms Tumor Protein primary antibodies -
Description
Rabbit monoclonal [EPR23963-116] to Wilms Tumor Protein -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC-Fr or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, K-562 and NIH:OVCAR-3 whole cell lysates; Mouse and rat testis tissue lysate. IHC-P: Human kidney, testis and ovarian carcinoma tissue; Mouse testis tissue; Rat testis tissue. ICC/IF: OVCAR-3 cells.
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23963-116 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab267377 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 52, 54 kDa (predicted molecular weight: 49 kDa).
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| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| ICC/IF |
1/50.
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| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 52, 54 kDa (predicted molecular weight: 49 kDa). |
|
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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ICC/IF
1/50. |
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
- Entrez Gene: 24883 Rat
- Omim: 607102 Human
- SwissProt: P19544 Human
- SwissProt: P22561 Mouse
- SwissProt: P49952 Rat
- Unigene: 591980 Human
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Alternative names
- WIT 2 antibody
- WT 1 antibody
- AWT1 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized OVCAR-3 (human ovary adenocarcinoma epithelial cell) cells labelling Wilms tumor protein with ab267377 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in OVCAR-3 cells. Negative control: LnCap (PMID:11299720). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-1-anti-wilms-tumor-protein-antibody-epr23963116-immunohistochemistry-human-kidney.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on human renal glomeruli (PMID:27922671). The section was incubated with ab267377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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![Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-463706-anti-wilms-tumor-protein-antibody-epr23963-116-western-blot-wildtype-hap1-wt1-knockout-hap1-k562-shsy5y.jpg)
Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)All lanes : Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : Wt1 knockout HAP1 cell lysate
Lane 3 : K-562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate
Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 50-60 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab267377 observed at 50-60 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab267377 was shown to react with Wilms Tumor Protein in wild-type HAP1 cells in Western blot with loss of signal observed in Wt1 knockout sample. Wild-type HAP1 and Wt1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab267377 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-412575-anti-wilms-tumor-protein-antibody-epr23963116-western-blot-myelogenous.jpg)
Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)All lanes : Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377) at 1/1000 dilution
Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 49 kDa
Observed band size: 52,54 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 10438524).
Negative control: LNCaP (PMID:11299720).
Exposure time: 26 seconds.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-4-anti-wilms-tumor-protein-antibody-epr23963116-immunohistochemistry-mouse-testis.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on Sertoli cells of mouse testis (PMID:17229929). The section was incubated with ab267377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
![Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-412578-anti-wilms-tumor-protein-antibody-epr23963116-western-blot-lysate.jpg)
Western blot - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)All lanes : Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377) at 1/1000 dilution
Lane 1 : Mouse testis tissue lysate
Lane 2 : Rat testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 49 kDa
Observed band size: 52,54 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 10438524).
Exposure time: 3 minutes.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-2-anti-wilms-tumor-protein-antibody-epr23963116-immunohistochemistry-human-testis.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on Sertoli cells of human testis (PMID:17229929). The section was incubated with ab267377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-3-anti-wilms-tumor-protein-antibody-epr23963116-immunohistochemistry-human-ovarian-carcinoma.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on human ovarian carcinoma (PMID:16547468). The section was incubated with ab267377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-5-anti-wilms-tumor-protein-antibody-epr23963116-immunohistochemistry-rat-testis.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Wilms tumor protein with ab267377 at 1/500 dilution followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining on Sertoli cells of rat testis (PMID:17229929). The section was incubated with ab267377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
![Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)](https://www.abcam.com/ps/products/267/ab267377/Images/ab267377-9-benefits-of-recombinant-antibodies.png)
Anti-Wilms Tumor Protein antibody [EPR23963-116] (ab267377)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab267377 has been referenced in 1 publication.
- Zhao J et al. Bruton's tyrosine kinase regulates macrophage-induced inflammation in the diabetic kidney via NLRP3 inflammasome activation. Int J Mol Med 48:N/A (2021). PubMed: 34278465