Thermo Scientific HCT 116 (+hIGF-1) Control Lysate is a stimulated cell lysate that has been validated for the expression levels and phosphorylation of key targets in the RAS signaling pathway.
HCT 116 (+hIGF-1) Control Lysate features include:
• Validated for the expression RAS by western blotting and of NRAS, KRAS, and HRAS by mass spectrometry analysis
• Optimized—stimulation and lysis conditions optimized for detection of RAS signaling targets
• Convenient—lysate is ready-to-use for immunoprecipitation, immunoprecipitation to mass spectrometry, and western blotting applications
• Stable for two years at -80°C
The RAS family of GTPase proteins modulates two key signaling pathways involved in cell growth and proliferation: the MAPK and PI3K pathways. Aberrant activation of RAS proteins promotes constitutive activation of cell growth and proliferation that can result in cancer. Mutations in the NRAS, KRAS, and HRAS oncogenes are the most common mutations in human cancer. This lysate has been validated for expression levels of the three isoforms of RAS using mass spectrometry.
The HCT 116 cell line is a human colon cancer epithelial cell line and has been used for proliferation and tumorigenicity studies for colon cancer. The HCT 116 (+hIGF-1) cell lysate is provided as frozen lysate at a standard concentration of 1 mg/mL (500 mg per vial). Lysates are prepared in non-denaturing buffer without loading dyes or dissociating agents (i.e., no 2-mercaptoethanol or dithiothreitol has been added). For western blot analysis, heat lysate to 95°C for 5 minutes. Add reducing agent before heating if desired. The recommended loading volume per lane is 10–20 μL (10–20 μg).
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