Human Gas6 Antibody

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.

Detection of Human Gas6 by Western Blot Migration of H1299 NSCLC cells enhanced by ligand-dependent Axl activation. (A) Western blotting to assess Gas6 expression in H1299 cells. Expression of Gas6 in LCAFhTERT cells was used as a positive control. (B) Phosphorylation of Axl was analyzed by Western blotting of whole cell lysates using different antibodies. GAPDH was used as an internal control. H1299 cells were stimulated for 15 min with 400 nM recombinant human Gas6 (rGas6). H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. (C) Migration of H1299 cells was analyzed using rGas6 (400 nM) added to the lower chamber. H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. (D) Western blotting of conditioned medium from LCAFhTERT transfected with siGas6 or siScr (control) to assess whether they contains Gas6 secreted by CAFs. (E) Phosphorylation of Axl in H1299 cells analyzed by Western blotting after stimulation with conditioned medium from siRNA-transfected LCAFhTERT. H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. The medium (DMEM containing 10% FBS) was used as control. (F) Migration of H1299 cells analyzed using conditioned medium of siRNA-transfected LCAFhTERT. H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. The medium (DMEM containing 10% FBS) was used as control. The relative number of migrated H1299 cells is indicated on the y-axis. Data show the mean ± SEM (n = 3); **p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28878389), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Gas6 by Immunohistochemistry Expression of Axl and Gas6 in clinical samples. (A,B) Immunohistochemical analysis of Axl (A) or Gas6 (B) in non-small cell lung cancer tissues from patients who underwent surgery following preoperative chemotherapy or chemoradiotherapy. Insets show higher magnification of the areas indicated in the boxes. Scale bar, 50 μm. (A) Representative images showing tumor Axl-negative tumor tissues (left), and tumor Axl-positive tumor tissues (right). Tissues were stained with an anti-Axl antibody (brown) and counterstained with hematoxylin. (B) Representative images showing stromal Gas6-negative tumor tissues (left) and stromal Gas6-positive tumor tissues (right). Tissues were stained with an anti-Gas6 antibody (brown) and counterstained with hematoxylin. (C) Correlation between tumor Axl expression and stromal Gas6 expression in tumor tissues observed in (A and B). (D) Correlation between tumor Axl, stromal Gas6 expression and survival. Kaplan–Meier plot of disease-free survival in patients with non-small cell lung cancer who underwent surgery following preoperative chemotherapy or chemoradiotherapy, stratified according to tumor Axl and stromal Gas6 expression. Five-year disease-free survival in the patients with tumors expressing both tumor Axl and stromal Gas6 (n = 37) was significantly worse than in the both-negative group (n = 12) (21.9% vs 51.3%, p = 0.04). The five-year disease-free survival of tumor Axl-negative and stromal Gas6-positive group was 50.7%, and the difference in survival between this group and both-positive or both-negative group was not significant (p = 0.20 and 0.49, respectively); *p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28878389), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Gas6 by Western Blot Migration of H1299 NSCLC cells enhanced by ligand-dependent Axl activation. (A) Western blotting to assess Gas6 expression in H1299 cells. Expression of Gas6 in LCAFhTERT cells was used as a positive control. (B) Phosphorylation of Axl was analyzed by Western blotting of whole cell lysates using different antibodies. GAPDH was used as an internal control. H1299 cells were stimulated for 15 min with 400 nM recombinant human Gas6 (rGas6). H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. (C) Migration of H1299 cells was analyzed using rGas6 (400 nM) added to the lower chamber. H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. (D) Western blotting of conditioned medium from LCAFhTERT transfected with siGas6 or siScr (control) to assess whether they contains Gas6 secreted by CAFs. (E) Phosphorylation of Axl in H1299 cells analyzed by Western blotting after stimulation with conditioned medium from siRNA-transfected LCAFhTERT. H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. The medium (DMEM containing 10% FBS) was used as control. (F) Migration of H1299 cells analyzed using conditioned medium of siRNA-transfected LCAFhTERT. H1299 cells were treated with or without TP-0903 (0.2 µmol/L) for 24 h. The medium (DMEM containing 10% FBS) was used as control. The relative number of migrated H1299 cells is indicated on the y-axis. Data show the mean ± SEM (n = 3); **p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28878389), licensed under a CC-BY license. Not internally tested by R&D Systems.