ras activation elisa kit  (Cell Biolabs Inc)

Journal: Oncology Reports

Article Title: Rab25 suppresses colon cancer cell invasion through upregulating claudin‑7 expression

doi: 10.3892/or.2023.8685

Figure Lengend Snippet: Rab25 inactivates EGFR. Cancer cells were transfected with the indicated vectors for 24 h. (A) The cell lysates were analyzed by immunoblotting with the indicated antibodies. (B) The p-EGFR expression was visualized using immunofluorescence. Original magnification, ×400; scale bar, 20 µm. (C) Ras activity was analyzed using the Ras-GTP pull down assay. Total protein expressions were analyzed by immunoblotting with the indicated antibodies. (D) Ras activity was analyzed using the Ras activation ELISA kit (***P < 0.001 vs. control vector). (E) HCT-116 cells were transfected with the indicated vectors for 24 h. Invasion was analyzed by utilizing a 48-well chemotaxis chamber (**P < 0.01 vs. control vector, ***P < 0.001 vs. control vector and ### P < 0.001 vs. Rab25 overexpression). Representative results of at least three independent experiments with similar results. Rab25, Ras-related protein 25; p-, phosphorylated; IP, immunoprecipitation; WB, western blotting.

Article Snippet: The cells were cultured to ~80% confluency in a 35 mm dish and were transfected for 24 h. The supernatants were removed and RAS activation was determined using a RAS activation ELISA kit (cat. no. 17-497; Millipore Sigma) according to the manufacturer's instructions.

Techniques: Transfection, Western Blot, Expressing, Immunofluorescence, Activity Assay, Pull Down Assay, Activation Assay, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation, Chemotaxis Assay, Over Expression, Immunoprecipitation