protein digestion
- Simultaneously confirm trypsin activity and protein digestion without performing gel electrophoresis.
- Accurately quantify % protein digested.
Protein identification by mass spectrometry is one of the most important steps in proteomics. Proteins are typically digested into peptides prior to the mass spectrometry (MS) step. The most popular enzyme used for protein digestion is trypsin. This enzyme is particularly suited for digestion because it has a very well defined specificity. Currently, MS and proteomics workflow processes do not include a step to verify that the trypsin is active and/or that the target protein has been adequately digested. Thus, tryptic digests are analyzed by MS without knowing whether the protein was digested or not.
ITSIPrep Protein Digestion Monitoring (ProDM, K-0021-10) kit (Patent pending) is a distinctive product that allows precise determination of the % protein digested using a proprietary colorimetric reagent and any spectrophotometer. By using the protein standard provided in the kit, you are also able to simultaneously determine if your enzyme (e.g. trypsin, chymotrypsin) is active. ProDM kit eliminates the need to perform gel electrophoresis, to confirm that the enzyme is active and the protein is adequately digested. ProDM will reduce the number of failed mass spectrometry experiments attributable to inactive enzyme and/or inadequate protein digestion, and hence the number of failed experiments.
Materials provided in the kit:
- Protein Standard
- Standard Buffer
- Reaction Buffer
- Reaction Quencher
- Detection Reagent
- Microfuge tubes
- Validated Procedure
 Use of ProDM kit to monitor the digestion of Bovine Serum Albumin (BSA) by trypsin. Panel "A" shows the spectrophotometric readings at various time intervals. Absorbance of the reaction mixture was measured at 585nm after addition of ITSIPREP Digestion Detection Reagent. Panel "B" shows electrophoretic image of the corresponding aliquots. Note that the BSA protein band in Panel B is only visible at T0 (no digestion). SM is size marker
|
HOME > Protein Research > Protein digestion monitoring
