|Detection of Human ZAG by Western Blot. Western blot shows lysates of human prostate tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human ZAG Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4764) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for ZAG at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.|
|ZAG in Human Breast Cancer Tissue. ZAG was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Goat Anti-Human ZAG Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4764) at 10 µg/mL 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 using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
|ZAG in Human Mesenchymal Stem Cells. ZAG was detected in immersion fixed human mesenchymal stem cells differentiated into adipocytes using Goat Anti-Human ZAG Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4764) at 10 µg/mL for 3 hours at room temperature. Cells were differentiated using using StemXVivo Osteogenic/Adipogenic Base Media (Catalog # CCM007) and StemXVivo Adipogenic Supplement (100X) (Catalog # CCM011). Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.|
ZAG (zinc-alpha 2-glycoprotein; also ZA2G) is a 40 kDa, secreted member of the MHC class I family of proteins. It is produced by adipocytes and various epithelial cells that generate exocrine-type secretions. ZAG is reported to stimulate lipid breakdown and thus may play a role in lipid homeostasis. Mature human ZAG is 278 amino acids (aa) in length. It contains one MHC class I antigen region (aa 26‑201) and a C2‑type Ig‑like domain (aa 207‑292). Two alternate splice forms exist; one shows a 66 aa substitution for the C‑terminal 30 aa, and a second shows a nine Lys substitution for aa 151‑298. Mature human ZAG is 60% aa identical to mouse ZAG.