MeCAT Duplex labeling kit for proteomics
The MeCAT Duplex kit is used for differential labeling of proteins and protein mixtures of two samples for differential and absolute quantification of their protein profiles (proteomes). The MeCAT kit consists of the MeCAT reagents for 2x8 reactions, labelling buffer and components. Two different MeCAT metal tags (Duplex kit: 8x a light lanthanide and 8x a heavy lanthanide, in total 16 reactions) are provided sufficient for 50-100 µg protein/reaction.
A four-plex kit containing Terbium, Holmium, Thulium and Lutetium-coded MeCAT reagents with 4x8 reactions is available upon request.
General MeCAT Labeling Workflow
- Protein extraction of two samples which should be compared
- Labelling of two protein samples with light and heavy MeCAT metal reagent
- Pooling of the two samples labelled with light and heavy MeCAT reagent
- Separation of MeCAT labeled proteins by 2DE, SDS-PAGE or chromatography
- (Absolute) quantification by ICP-MS
- Identification by ESI-MS or MALDI-MS
Example MeCAT Workflow Ultra sensitive absolute and relative MeCAT protein quantification is performed by ICP-MS (inductively coupled plasma mass spectrometry) on protein level down to low attomol range. Proteins can be identified and relatively quantified on peptide level by MALDI-MS an ESI-MSMS. Metal standard solutions are used for high accuracy external protein independ calibration of ICP-MS.