Cells were homogenized in RIPA buffer as described17 (link). Nuclear and cytoplasmic extracts were isolated by centrifugation and hypotonic lysis as described18 (link). Antibodies for analyses included: RON (SC-322), Androgen Receptor, (SC-815), Axl (SC-1096), Gas6 (SC-1935), and Tubulin (SC-5286) from Santa Cruz Biotechnology; Src (2110S), phosphor-Src y416 (2101S), Akt (4691S), phosphor-Akt s473 (4060S), phospho-Axl y702 (5724S) and LAMIN A/C (4777S) from Cell Signaling Technologies; phospho-RON y1238/y1239 (AF1947, R&D); ACTIN (Cincinnati Children’s Hospital Medical Center, Clone C4). Peroxidase-conjugated secondary antibodies (Jackson Laboratories) were applied, and membranes were developed using Pierce ECL2 Western Blotting substrate (ThermoFisher Scientific). Membranes were stripped using Restore Western Blot Stripping Buffer (ThermoFisher Scientific) before re-probing. The Proteome Profiler Mouse Phospho-RTK Array Kit (ARY014, R&D) was used on whole tumor lysates and performed according to manufacturer’s instructions.
Phospho ron y1238 y1239 af1947
Manufactured by R&D Systems
Phospho-RON y1238/y1239 (AF1947) is a primary antibody used to detect phosphorylation of the RON receptor tyrosine kinase at tyrosine residues 1238 and 1239.
Automatically generated - may contain errors
Lab products found in correlation
Peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions)
Tubulin sc 5286, by Santa Cruz Biotechnology (2 mentions)
Proteome profiler mouse phospho rtk array kit, by R&D Systems (2 mentions)
Lamin a c 4777, by Cell Signaling Technology (2 mentions)
Restore western blot stripping buffer, by Thermo Fisher Scientific (2 mentions)
Pierce ecl 2 western blotting substrate, by Thermo Fisher Scientific (2 mentions)
2 protocols using phospho ron y1238 y1239 af1947
1
Protein Extraction and Western Blot Analysis
2
Western Blot Analysis of Signaling Pathways
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Cells were homogenized in RIPA buffer as described.17 Nuclear and cytoplasmic extracts were isolated by centrifugation and hypotonic lysis as described.18 Antibodies for analyses included: RON (SC‐322), Androgen Receptor, (SC‐815), Axl (SC‐1096), Gas6 (SC‐1935), and Tubulin (SC‐5286) from Santa Cruz Biotechnology; Src (2110 S), phosphor‐Src y416 (2101 S), Akt (4691 S), phosphor‐Akt s473 (4060S), phospho‐Axl y702 (5724S), and LAMIN A/C (4777S) from Cell Signaling Technologies; phospho‐RON y1238/y1239 (AF1947, R&D); ACTIN (Cincinnati Children's Hospital Medical Center, Clone C4). Peroxidase‐conjugated secondary antibodies (Jackson Laboratories) were applied, and membranes were developed using Pierce ECL2 Western Blotting substrate (ThermoFisher Scientific). Membranes were stripped using Restore Western Blot Stripping Buffer (ThermoFisher Scientific) before re‐probing. The Proteome Profiler Mouse Phospho‐RTK Array Kit (ARY014, R&D) was used on whole tumor lysates and performed according to the manufacturer's instructions.
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