Human HGFR/c-MET Antibody

HGF R/c‑MET in HT‑29 Human Cell Line.

HGF R/c-MET was detected in immersion fixed HT-29 human colon adenocarcinoma cell line using Mouse Anti-Human HGF R/c-MET Monoclonal Antibody (Catalog # MAB3581) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane and secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HGF R/c‑MET in Human Renal Cell Carcinoma Tissue.

HGF R/c-MET was detected in immersion fixed paraffin-embedded sections of human renal cell carcinoma tissue using Mouse Anti-Human HGF R/c-MET Monoclonal Antibody (Catalog # MAB3581) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). 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 DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human HGFR/c-MET by ELISA

ABT-700 specifically binds cellular c-Met and antagonizes c-Met signaling in both HGF-dependent and -independent settings. a FACS analysis of ABT-700 binding to MCF7 transfectants. Stable human c-Met or vector control transfectants of human MCF7 breast cancer cells were incubated with increasing amounts of ABT-700 and bound ABT-700 was detected by FACS with secondary anti-human IgG conjugated with Alexa 488. b ELISA quantification of phospho-c-Met in A549 cells. A549 cells grown in a 96-well plate were pre-incubated for one hour with antibodies in a dose-range as shown, followed by stimulation with 1 nM HGF for 10 min. Total cell lysates were made and phospho-c-Met was detected by ELISA. c ELISA quantification of phospho-c-Met in SNU5 cells. SNU5 cells grown in a 96-well plate were incubated with antibodies in a dose-range as shown for 6 h. Total cell lysates were made and subjected to ELISA for phospho-c-Met. The value of cells in media alone was used as 100 % of control. d ELISA quantification of total c-Met in SNU5 cells. SNU5 cells grown in a 96-well plate were incubated with antibodies in a dose-range as shown for 6 h. Total cell lysates were made and c-Met level was determined by ELISA. The value of cells in media alone was used as 100 % of control. e Western blot analysis of U87MG cell lysates. U87MG cells grown in a 12-well plate were treated with antibodies as shown at 10 μg/mL for 10 min, 1 h or 6 h. Total cell lysates were analyzed for c-Met and other phosphorylated targets as shown. Western blot analysis of Hs746T cell lysates. Hs746T cells grown in a 12-well pate were treated with antibodies as shown at 10 μg/mL for 6 h. Total cell lysates were analyzed for c-Met and other phosphorylated targets as shown. f Western blot analysis of SNU620 cell lysates. SNU620 cells grown in a 12-well pate were treated with antibodies as shown at 10 μg/mL for 24 h. Total cell lysates were analyzed for c-Met and other phosphorylated targets as shown. g Inhibition of proliferation of SNU620 cells. SNU620 cells were plated in a 96-well plate and treated with antibodies in a dose range as shown for 3 days. Quantification of live cells at the end of incubation was done with Cell-titer Glo reagents. The data shown in all panels are from one of at least two independent experiments showing similar results as described in Methods Image collected and cropped by CiteAb from the following publication (https://www.biomedcentral.com/1471-2407/16/105), licensed under a CC-BY license. Not internally tested by R&D Systems.