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 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
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.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
A novel source of cultured podocytes. Authors: Da Sacco, Stefano, Lemley, Kevin V, Sedrakyan, Sargis, Zanusso, Ilenia, Petrosyan, Astgik, Peti-Peterdi, Janos, Burford, James, De Filippo, Roger E, Perin, Laura PLoS ONE, 2013;8(12):e81812. Species: Human Sample Type: Whole Cells Application: Flow
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