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.
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.
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.
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.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
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.
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.
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