Detection of Human Gas6 by Western Blot.
Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line, SW480 human colorectal adenocarcinoma cell line, COLO 205 human colorectal adenocarcinoma cell line, and human intestine tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Gas6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF885) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for Gas6 at approximately 64 and 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Gas6 in Human Stomach Cancer Tissue.
Gas6 was detected in immersion fixed paraffin-embedded sections of human stomach cancer tissue using 10 µg/mL Goat Anti-Human Gas6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF885) 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). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
Gas6 (Growth Arrest Specific 6) is a multimodular protein that is upregulated by a wide variety of cell types in response to growth arrest (1). Gas6 and the structurally related Protein S are vitamin K-dependent and have an extensively gamma -carboxylated N-terminal Gla domain, four EGF-like repeats, and a C-terminal region with homology to steroid hormone binding globulin (SHBG) (2). Human Gas6 is a 75 kDa protein that shares 77-79% amino acid (aa) sequence identity with mouse and rat Gas6 and 43% aa sequence identity with human protein S (over the region expressed). Alternate splicing generates isoforms that lack the Gla domain and/or the spacer between the EGF-like and SHBG regions. Gas6 binds and induces signaling through the receptor tyrosine kinases Axl, Dtk, and Mer (3-5). Human Gas6 interacts with both mouse and rat orthologs of these receptors (1). The full length isoform may be cleaved, resulting in release of the free SHBG region which can independently activate Axl (6). Shed soluble forms of Axl and Mer bind Gas6 and function as decoy receptors (7, 8). Gas6 induces a variety of responses, including prevention of apoptosis (9), cell proliferation (10), platelet-mediated thrombosis (11), retinal epithelial cell phagocytosis of outer rod segments (12), inhibition of VEGF‑induced endothelial cell chemotaxis (13), and the differentiation and expansion of NK cell precursors (14). The affinity of Gas6 for phosphatidylserine likely contributes to its role in promoting the phagocytosis of apoptotic cells (15). Several of these effects have been shown to require gamma -carboxylation of the Gla domain (12, 16).
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