ProteoSpin™ On-Column Proteolytic Digestion Kit
For the rapid and convenient on-column digestion of proteins
For research use only and NOT intended for in vitro diagnostics.
ProteoSpin™ On-Column Proteolytic Digestion Kit
For the rapid and convenient on-column digestion of proteins
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Overview
- Rapid and simple procedure to generate digested peptides
- Simultaneous digestion, purification and concentration at once
- Peptide generation is complete, with no generation of additional artifacts being detected in mass spectrometry
- Peptides are ready for applications such as mass spectrometry and SDS-PAGE
- Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
This kit is highly efficient in the enzymatic digestion of simple and complex protein samples using trypsin and the subsequent purification of the resulting peptides using a convenient spin column format. Trypsin is added to the protein sample and bound to the column. Salts are washed away and the trypsin is then activated to digest proteins. Peptides are then eluted in a small volume and ready for downstream analysis. The peptides generated are complete, with no additional artifacts being detected in mass spectrometry. Fifteen micrograms of protein can be processed, digested and purified with each spin column with about 20 minutes of hands-on time (plus trypsin incubation). The simultaneous protein digestion and volumetric concentration of the purified peptides makes the kit a convenient method for preparing peptides to be analyzed by many downstream applications such as mass spectrometry and more.
Details
Supporting Data
Figure 1. Generation of Tryptic BSA Peptides without the Generation of any Artifacts. MALDI-TOF spectra of the peptides that resulted after BSA was digested on-column with trypsin using the ProteoSpinTM On-Column Proteolytic Digestion Kit (Panel A) or digested in a typical liquid environment (Panel B). For both digestions, 15 µg of BSA was used as the input and 0.3 µg of Trypsin was used for the digestion. For on-column digestion, the 15 µg of BSA was loaded onto the column with the trypsin, the BSA was washed, then the on-column digestion proceeded and the resulting peptides were eluted. By comparing the spectra, it can be seen that the ProteoSpinTM kit (Panel A) resulted in the generation of the same peptide profile in MALDI-TOF as the liquid digestion did (Panel B). Furthermore, no artifacts were found to arise from the use of the column, and all the same peaks could be detected in both spectra.
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Kit Specifications
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| Maximum Protein Input |
15 μg
|
| Minimum Protein Input |
2 μg
|
| Minimum Elution Volume |
30 μL
|
| Time to Process 10 Purifications |
20 minutes
(Plus a 1-hour incubation) |
Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 17500 (25 preps) |
|---|---|
| Wash Solution C | 30 mL |
| Binding Buffer A | 4 mL |
| Column Activation Buffer | 3 mL |
| Micro Spin Columns | 25 |
| Collection Tubes | 25 |
| Elution tubes (1.7 mL) | 25 |
| Product Insert | 1 |
Documentation
Citations
| Title | ADNP dysregulates methylation and mitochondrial gene expression in the cerebellum of a Helsmoortel-Van der Aa syndrome autopsy case |
| Citation | Acta Neuropathologica Communications 2023. |
| Authors | Claudio D'Incal, Anke Van Dijck, Joe Ibrahim, Kevin De Man, Lina Bastini, Anthony Konings, Ellen Elinck, lllana Gozes, Zlatko Marusic, Mirna Anicic, Jurica Vukovic, Nathalie Van der Aa, Ligia Mateiu, Wim Vanden Berghe & R. Frank Kooy |