TUBE-based Mass Spectrometry Proteomics
As a LifeSensors customer, your proteomics project begins when our team receives your sample as a frozen cell pellet. Our required size is a minimum of 2.5x108 cells / 5 mg protein. We recommend a pellet size of, 5-10x108 cells / 10-20 mg protein. Once your tray is received, we perform cell lysis, TUBE enrichment of polyubiquitylated proteins, in-gel trypsin digestion of enriched sample, and LC-MS/MS analysis.
Upon completion of the analysis, you will receive a list of peptides and corresponding proteins which have been identified. This information will help you publish your results or plan your next assay.
Each experiment is unique, therefore the time required for individual experiments may differ. However, we do provide a rough estimate based upon the number of samples you want analyzed:
3-5 week turnaround for 1-4 samples
4-6 weeks for 5-9 samples
5-7 weeks for 10-15 samples
Please contact us for time estimates for analysis of 16 or more samples.
TUBE-based enrichment of polyubiquitinated proteins has proven key for the progress of ubiquitin proteomics. LifeSensors have developed two proprietary technologies to- (1) identify cellular proteins that are ubiquitinated (TUBE proteomics), and (2) qualitative and quantitative profiling of ubiquitination sites on proteins (UbiSight proteomics). A brief overview of these two technologies are depicted in the following figure.
LifeSensors’ pan (TUBE1 and TUBE2) as well as linkage selective (M1-, K48-, and K63) TUBEs provide isolation of respective polyubiquitinated proteins. The TUBE enrichment kit contains solutions to provide simple procedure to lyse cells, wash the non-specific binding proteins, and elute proteins in solution compatible for further protease digestion. TUBE proteomics services include following four steps:
- TUBE Enrichment Kit - Kit for enrichment of polyubiquitinated protein: Magnetic-TUBE1 and TUBE2 beads, Cell lysis buffer, Wash buffer and Elution buffer
- K48 TUBE Enrichment Kit - Kit for enrichment of K48-polyubiquitinated proteins: K48-linkage selective TUBE, Cell lysis buffer, Wash buffer and Elution buffer
- K63 TUBE Enrichment Kit - Kit for enrichment of K63-polyubiquitinated proteins: K63-linkage selective TUBE, Cell lysis buffer, Wash buffer and Elution buffer
- M1 TUBE Enrichment Kit- Kit for enrichment of M1 (linear)-polyubiquitinated proteins: M1-linkage selective TUBE, Cell lysis buffer, Wash buffer and Elution buffer