WT1 Antibody [Unconjugated]

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.

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.

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 (//pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//www.nature.com/articles/srep08826), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/30222766), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//www.nature.com/articles/srep08826), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

WT1 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human WT1
  Met127-Gly249
  Accession # P19544
• Specificity: Detects human WT1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: WT1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for WT1 Antibody [Unconjugated]

• AWT1
• GUD
• NPHS4GUD
• WAGR
• Wilms tumor 1
• Wilms tumor protein
• WIT-2
• WT1
• WT33

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for WT1 Antibody (AF5729)(5)

Publications using AF5729

• Abdollahzadeh, F;Khoshdel-Rad, N;Bahrehbar, K;Erfanian, S;Ezzatizadeh, V;Totonchi, M;Moghadasali, R; Enhancing maturity in 3D kidney micro-tissues through clonogenic cell combinations and endothelial integration Journal of cellular and molecular medicine 2024-06-01 [PMID: 38818569] (Immunohistochemistry, Mouse)
• Whitehead AJ, Hocker JD, Ren B, Engler AJ Improved epicardial cardiac fibroblast generation from iPSCs Journal of molecular and cellular cardiology 11/24/2021 [PMID: 34826415]
• C Rauch, E Feifel, G Kern, C Murphy, F Meier, W Parson, M Beilmann, P Jennings, G Gstrauntha, A Wilmes Differentiation of human iPSCs into functional podocytes PLoS ONE, 2018-09-17;13(9):e0203869. 2018-09-17 [PMID: 30222766] (Western Blot, Human)
• Imberti , Barbara, Tomasoni , Susanna, Ciampi , Osele, Pezzotta , Anna, Derosas , Manuela, Xinaris , Christod, Rizzo , Paola, Papadimou , Evangeli, Novelli , Rubina, Benigni , Ariela, Remuzzi , Giuseppe, Morigi , Marina Renal progenitors derived from human iPSCs engraft and restore function in a mouse model of acute kidney injury. Sci Rep, 2015-03-06;5(0):8826. 2015-03-06 [PMID: 25744951] (IHC, Human)
• Da Sacco , Stefano, Lemley , Kevin V, Sedrakyan , Sargis, Zanusso , Ilenia, Petrosyan , Astgik, Peti-Peterdi , Janos, Burford , James, De Filippo , Roger E, Perin , Laura A novel source of cultured podocytes. PLoS ONE, 2013-12-12;8(12):e81812. 2013-12-12 [PMID: 24349133] (Flow Cytometry, Human, Mouse)

Bioinformatics

• Gene Symbol: WT1
• Uniprot:
  • P19544(Human)