Intracellular Staining by Flow Cytometry
|Detection of Patched 2 in WS‑1 Human Cell Line by Flow Cytometry. WS‑1 human fetal skin fibroblast cell line was stained with Human Patched 2 (First Extracellular Loop) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF4078, filled histogram) or isotype 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.|
|Patched 2 in Human Basal Cell Carcinoma. Patched 2 was detected in immersion fixed paraffin-embedded sections of human basal cell carcinoma using 3 µg/mL Goat Anti-Human Patched 2 (First Extracellular Loop) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4078) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of keratinocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
Patched 2 (Ptch2) is a 180-200 kDa 12 transmembrane member of the Patched family. Human PTCH2 is 1203 amino acids (aa) in length. It has two terminal cytoplasmic domains, one between aa 1-57, and a second between aa 1115-1203. Together with Smoothened (SMO), Ptch2 forms a membrane complex that serves as a receptor for Sonic Hedgehog (SHH). Human Ptch2 likely has two isoforms. One shows an eight aa extension of the C-terminus, while a second shows a four aa substitution for the C-terminal 61 amino acids (aa 1143-1203). Over the first extracellular loop human Patched 2 shares 95% and 96% aa sequence identity with mouse and canine Patched 2, respectively.
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.
The document you requested is not available online. Please enter the Catalog Number and Lot Number below to have a document emailed to you at the address provided