Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all WT1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human WT1
Met127-Gly249
Accession # P19544

Specificity

Detects human WT1 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human WT1 Antibody

Detection of Human WT1 antibody by Western Blot.

Detection of Human WT1 by Western Blot.

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human WT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5729) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for WT1 at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of WT1 antibody in HL‑60 Human Cell Line antibody by Flow Cytometry.

Detection of WT1 in HL‑60 Human Cell Line by Flow Cytometry.

HL-60 human acute promyelocytic leukemia cell line was stained with Goat Anti-Human WT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5729, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Immunostaining of podocyte-like cells derived from the iPSC line SBAD3 with podocyte markers and F-actin staining.iPSC were differentiated on glass cover slips fixed and stained for synaptopodin, WT-1, podocin, and F-actin as described in methods. Red and green colours were applied post capture. Staining in two other iPSC lines are provided in S1 and S2 Figs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Immunostaining of podocyte-like cells derived from the iPSC line SBAD3 with podocyte markers and F-actin staining.iPSC were differentiated on glass cover slips fixed and stained for synaptopodin, WT-1, podocin, and F-actin as described in methods. Red and green colours were applied post capture. Staining in two other iPSC lines are provided in S1 and S2 Figs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human WT1 by Western Blot

Detection of Human WT1 by Western Blot

Immunostaining of podocyte-like cells derived from the iPSC line SBAD3 with podocyte markers and F-actin staining.iPSC were differentiated on glass cover slips fixed and stained for synaptopodin, WT-1, podocin, and F-actin as described in methods. Red and green colours were applied post capture. Staining in two other iPSC lines are provided in S1 and S2 Figs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Immunostaining of podocyte-like cells derived from the iPSC line SBAD3 with podocyte markers and F-actin staining.iPSC were differentiated on glass cover slips fixed and stained for synaptopodin, WT-1, podocin, and F-actin as described in methods. Red and green colours were applied post capture. Staining in two other iPSC lines are provided in S1 and S2 Figs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Differentiation of human iPSC clone IV towards renal commitment.(a) Representative immunofluorescence images of co-staining for Lhx1/Osr1 up to day 6. (b) Images of co-staining of Pax2/Six2, Pax8/Six2 and Six2/Wt1 from day 6 to 12. (c) Expression of renal progenitor markers such as CD24, Claudin1 and GGT1 from day 0 to 19. Nuclei are stained with DAPI (blue). Scale bars: 50 μm (a, b), 20 μm (c). (d) Gene expression analysis for renal progenitor markers at different points in time. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08826), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Immunostaining of podocyte-like cells derived from the iPSC line SBAD3 with podocyte markers and F-actin staining.iPSC were differentiated on glass cover slips fixed and stained for synaptopodin, WT-1, podocin, and F-actin as described in methods. Red and green colours were applied post capture. Staining in two other iPSC lines are provided in S1 and S2 Figs. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Detection of Human WT1 by Immunocytochemistry/Immunofluorescence

Stepwise differentiation of human iPSCs towards renal progenitor cells (RPCs).(a) Schematic description of the two-stage protocol applied to iPSC-renal commitment. (b, c, d, e) Immunofluorescence of iPSCs (derived from retroviral transfected dermal fibroblasts) exposed to differentiating media. (b) The pluripotency markers SSEA4, TRA-1-81, Nanog, ME marker T(Bry) and IM marker Osr1. (c) IM and MM marker expression as Wt1, Pax8, Pax2, Six2 and Sall1. (d) Renal progenitor markers CD133, CD24, NCAM, glomerular epithelial marker Claudin1 and proximal tubular epithelial markers AQP1, GGT1. (e) Markers identifying endodermal AFP, ectodermal Pax6, and cardiac mesodermal Nkx2.5 derivation. Nuclei are stained with DAPI (blue). Scale bars: 20 μm (b, c, d, e). Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/srep08826), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human WT1 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HL‑60 human acute promyelocytic leukemia cell line fixed with paraformaldehyde and permeabilized with saponin

Western Blot

1 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: WT1

WT1 (Wilms’ tumor protein 1; also WT33) is a 52‑54 kDa, nuclear member of the EGR C2H2‑type zinc‑finger family of proteins. Although its predicted MW is 49 kDa, it runs anomalously in SDS‑PAGE, likely due to a high proline content. It is widely expressed, being found in developing Sertoli cells, glomerular podocytes, neurons, and CD34+ stem cells. Human WT1 is 449 amino acids (aa) in length. It contains a Pro‑rich domain (aa 27‑83) and four consecutive C2H2 zinc finger regions
(aa 323‑347; 353‑377; 383‑405; 414‑438). WT1 forms homodimers, and interacts with multiple molecules. Interaction with the zinc fingers generally promotes gene transcription, while N‑terminal interactions block gene transcription. There are at least two dozen splice variants. Some are combinations of deletions of aa 250‑266 and 408‑410, plus an alternate start site 68 aa upstream of the standard site, and a three aa substitution for aa 1‑147. Over aa 127‑249, human WT1 shares 98% aa identity with mouse WT1.

Long Name

Wilms Tumor 1

Alternate Names

GUD, WAGR, WIT-2, WT33

Entrez Gene IDs

7490 (Human); 22431 (Mouse); 24883 (Rat)

Gene Symbol

WT1

UniProt

P19544 (Human)

Additional WT1 Products

Product Documents for Human WT1 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human WT1 Antibody

For research use only

Citations for Human WT1 Antibody

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