Integrating cell biology workflows into quantitative analytical platforms
Identify differences in diseased versus normal cells (A) and their role in signal transduction pathways (B) by comparing the relative abundance of individual proteins (C).
SILAC™ Protein ID and Quantitation Kits
Grow duplicate cultures in SILAC™ media with isotopically distinct amino acids (AA)
If needed apply stimulus (e.g. RNAi, etc.) to culture grown in SILAC™ Media. Harvest and mix cells.
Lyse cells using SILAC™ lysis buffers "run lysate on NuPAGE™ ID gel", excise gel band and trypsinize.
Sample submission to MS operator for data analysis.
Obtain list of proteins identified with relative abundances.