Cells were washed once with modified Dulbecco’s phosphate-buffered saline and lysed in buffer containing 0.025 M Tris, 0.15 M NaCl, 0.0001 M EDTA, 1% NP-40, and 5% glycerol (pH 7.4). Protein G/A agarose resin (Thermo Fisher Scientific, Waltham, MA) was incubated for 2 hours with rabbit monoclonal anti-MET antibody (Cell Signaling Technologies), anti-EGFR antibody (Cell Signaling Technologies) or rabbit IgG control (Invitrogen, Carlsbad, CA) as a negative control and cell lysate at 1 mg of protein was mixed with antibody-coupled resin (Cell Signaling Technologies) overnight at 4°C. Co-immunoprecipitation was performed using the Pierce Co-IP Kit (Thermo Fisher Scientific). Immunoprecipitates were washed, eluted, and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot was performed as described above.
Rabbit monoclonal anti met antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Rabbit monoclonal anti-MET antibody is a laboratory research tool used for the detection and analysis of the MET protein, also known as the hepatocyte growth factor receptor. This antibody is produced in rabbit and can be used for various immunological techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry.
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Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Anti egfr antibody, by Cell Signaling Technology (1 mentions)
Pierce co ip kit, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Fdbio (1 mentions)
Anti rabbit igg secondary antibody, by Abcam (1 mentions)
Anti α tubulin antibody, by Merck Group (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Alexa fluor 488 goat anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Fv1000, by Olympus (1 mentions)
3 protocols using rabbit monoclonal anti met antibody
1
Protein Co-Immunoprecipitation Assay
2
Western Blot Analysis of MET Protein
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Cells were lysed using RIPA buffer containing protease inhibitor cocktail (Fdbio Science, Hangzhou, China), and the protein concentrations were evaluated using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA). Total proteins were separated on 10% SDS-PAGE gels, transferred to polyvinylidene fluoride membranes (Merck Millipore, Billerica, MA, USA) and the membranes were incubated with rabbit monoclonal anti-MET antibody (1 : 1000; Cell Signaling Technology, Beverly, MA, USA), and then incubated with anti-rabbit IgG secondary antibody (1 : 5000; Epitomics, Burlingame, CA, USA). An anti-α-tubulin antibody (1 : 1000; Sigma-Aldrich) was used as the loading control and the bands were detected by enhanced chemiluminescence.
3
Immunofluorescence Microscopy of MET
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Transfected CNE-2 or SUNE-1 cells were seeded onto coverslips (Thermo Fisher Scientific), and 24 h later, the coverslips were fixed, permeabilized and incubated with rabbit monoclonal anti-MET antibody (1 : 3000; Cell Signaling Technology), and then incubated with the Alexa Fluor 488 goat anti-rabbit IgG secondary antibody (Life Technologies, Carlsbad, CA, USA). Later, the cells were counterstained with 4′, 6-diamidino-2-phenylindole (DAPI) and images were captured using a confocal laser scanning microscope (Olympus FV1000, Olympus, Tokyo, Japan).
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