Human Pygopus-2 Antibody Summary
Met160-Glu336
Accession # NP_612157
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human Pygopus‑2 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, MCF-7 human breast cancer cell line, and Daudi human Burkitt's lymphoma cell line. PVDF Membrane was probed with 0.5 µg/mL of Mouse Anti-Human Pygopus-2 Monoclonal Antibody (Catalog # MAB3616) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Pygopus-2 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Pygopus‑2 in MCF‑7 Human Cell Line. Pygopus-2 was detected in immersion fixed MCF-7 human breast cancer cell line using Mouse Anti-Human Pygopus-2 Monoclonal Antibody (Catalog # MAB3616) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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Detection of Human Pygopus‑2 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Pygopus‑2 at approximately 65 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Pygopus‑2 Monoclonal Antibody (Catalog # MAB3616). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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Pygopus‑2 in Human Ovary and Ovarian Cancer Tissue. Pygopus‑2 was detected in immersion fixed paraffin-embedded sections of normal human ovary (left panel) and ovarian cancer tissue (right panel) using Mouse Anti-Human Pygopus‑2 Monoclonal Antibody (Catalog # MAB3616) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Reconstitution Calculator
Preparation and Storage
- 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.
Background: Pygopus-2
Human Pygopus-2 (pygopus; the name of a legless Australian lizard) is a 41 kDa, 406 amino acid (aa) transcriptional coactivator. It contains three domains: an N‑terminal NLS (aa 41-47), followed by a proline-rich region and a zinc-finger PHD-type domain (aa 330-387). A potential internal alternate start site may generate an isoform that deletes the first 37 aa. Pygopus 2 is a nuclear protein that acts in concert with BCL-9 and TCF to retain beta -Catenin in the nucleus during Wnt-signaling. It does so by forming a “chain-of-adaptors” with the above three proteins. Over the range of aa used for immunization, human pygopus 2 is 95% identical to mouse.
Product Datasheets
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