The human colorectal cancer cell lines, HCT116, HT29 and DLD-1 and lung cancer cell line 226Br were cultured in RPMI1640 (Welgene, Seoul, Korea) containing 10% fetal bovine serum (FBS; Welgene) and antibiotics (100 mg/L penicillin and 100 mg/L streptomycin; GIBCO-BRL Life Technologies; Gaithersburg, MD, USA). Human colonic fibroblasts (CCD-18co) were cultured in DMEM (Welgene, Seoul, Korea) containing 10 % FBS and antibiotics Cells were incubated in a humidified thermostat under 5 % CO2 at 37 °C. A predesigned siRNA targeting HGF and scramble siRNA were purchased from Shanghai Genepharma (Shanghai, China). Recombinant Human HGF was purchased from R&D system (Minneapolis, MN, USA). Humanized anti-HGF antibody was kindly provided by YooYoung pharmaceutical Co. Ltd (Seoul, Korea) and iBIO Inc (Seoul, Korea). Irinotecan (CPT-11) was obtained from Sigma Aldrich (St. Louis, MO, USA). Antibodies to c-MET, phosphorylated-c-MET, phosphorylated-ERK, phosphorylated-AKT, PARP, and cleaved Caspase-3 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody against HGF, actin and fibroblast marker (ER-TR7) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Phosphorylated c met
Manufactured by Cell Signaling Technology
Sourced in United States
Phosphorylated-c-Met is a laboratory product that detects the phosphorylation status of the c-Met protein. c-Met is a receptor tyrosine kinase that plays a crucial role in various cellular processes. This product can be used to measure the activation and signaling of the c-Met pathway in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Phosphorylated akt, by Cell Signaling Technology (1 mentions)
Dulbecco modified eagle medium (dmem), by Welgene (1 mentions)
Irinotecan cpt 11, by Merck Group (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Recombinant human hgf, by R&D Systems (1 mentions)
Scramble sirna, by GenePharma (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Phosphorylated erk, by Cell Signaling Technology (1 mentions)
C met, by Cell Signaling Technology (1 mentions)
Rpmi 1640, by Welgene (1 mentions)
Phosphorylated c jun, by Cell Signaling Technology (1 mentions)
C met, by Merck Group (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Pvdf membrane, by GE Healthcare (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Bis tris gradient gel, by Thermo Fisher Scientific (1 mentions)
Blocker casein in pbs, by Thermo Fisher Scientific (1 mentions)
2 protocols using phosphorylated c met
1
Colorectal and Lung Cancer Cell Culture and Targeting HGF
2
Western Blot Analysis of Protein Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein extracted from tissues was subjected to electrophoresis using a 4–12% Bis-Tris gradient gel (Invitrogen, MA, United States) for 50 min. After transferring the gel to PVDF membranes (GE Healthcare, MA, United States), membranes were then blocked with Blocker Casein in PBS (Thermo Fisher, MA, United States) at room temperature (45 min), followed by the incubation with primary antibodies specific to c-Met (Sigma, MO, United States), phosphorylated c-Met (Cell Signaling Technology, MA, United States), c-Jun (Cell Signaling Technology, MA, United States), phosphorylated c-Jun (Cell Signaling Technology, MA, United States), and GAPDH (Cell Signaling Technology, MA, United States) 4 °C (overnight), respectively. Membranes were washed three times TBST (Invitrogen, MA, United States) and then subjected to incubation with a secondary antibody specific to anti-rabbit IgG (Cell Signaling Technology, MA, United States) at room temperature (1 h). Immuno-stained membranes were exposed to Super Signal West Femto Maximum Sensitivity Substrate (Thermo Fisher, MA, United States) or Super Signal West Pico Chemiluminescent Substrate (Thermo Fisher, MA, United States), and signal intensities were visualized using ImageQuant Las 4000 (GE Healthcare, MA, United States), followed by the quantification using ImageJ software (NIH).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!