Rabbit anti met antibody

For immunostaining of Met, frozen sections were fixed with methanol, tissue sections were incubated with PBS containing 3% bovine serum albumin for 30 min, washed with PBS, and incubated overnight with rabbit anti-Met antibody (Santa Cruz, code: sc-162, clone SP-260) in PBS containing 0.5% bovine serum albumin and 0.1% Tween-20 at 4 °C. After washing, sections were incubated with EnVision+ System-HRP Labeled Polymer (DAKO, code: K4002, Glostrup, Denmark). Immunocomplexes were visualized using an IMMPACT DAB kit (VECTOR, code: SK-4105, PE2 6XS, UK) and nuclei were stained with hematoxylin. For blood vessel staining, frozen sections were fixed with methanol, then treated with rat anti-CD31 (BD, code: 557355, clone MEC13.3) and mouse anti-α-smooth muscle actin (DAKO, code: M0851, clone 1A4, Glostrup, Denmark) antibodies were diluted via a Vector^® M.O.M.™ Immunodetection Kit (Vector Laboratories, Burlingame, CA). Sections were analyzed using a Biozero BZ-9000 (KEYENCE, Osaka, Japan).

Mouse anti-Abl antibody was from BD Bioscience. Mouse anti-Arg antibody, rabbit anti-CrkL antibody, rabbit anti-Met antibody, and rat anti-CSF-1R were from Santa Cruz Biotechnology. Phospho-specific antibodies for Abl (pY412) (BD) and Arg (pY439) (ABGENT) were used for western blotting. Mouse anti-tubulin antibody was from Sigma. Rabbit anti-phospho-CrkL (Y207) antibody, rabbit anti-Met (Y1234/1235) antibody, rabbit anti-Met (Y1349) antibody, rabbit anti-Met (Y1003) antibody, mouse anti-phospho-MLC2 (S19) antibody were from Cell Signaling Technology. Rat anti-E-cadherin was from Zymed (Invitrogen). Mouse anti-gp135 antibody was a gift from Dr. George Ojakian (SUNY Downstate Medical Center). Mouse anti-GFP antibody and Fugene 6 were from Roche. Hepatocyte growth factor (HGF) and GNF2 were from Sigma. pBabe-Sin-Puro-Tet-RhoA Biosensor (RhoA FRET reporter) was from Addgene. HGF used for 3D tubulogenesis was from ProSpec. OxyFluor used in live cell imaging was from Oxyrase Inc. Portein G sepharose was from GE healthcare. ProLong antifade mounting media was from Invitrogen.