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Application Note - Immobilized pepsin digestion of Cytochrome-C
ProDx columns
Download: Application Note - Immobilized pepsin digestion of Cytochrome-C [PDF]
Components
- Equine Cytochrome-C
Brochure - Automated Hydrogen-Deuterium Exchange
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Brochure - Automated Hydrogen-Deuterium Exchange (Japanese)
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Performance Report - ProDx Protease column
Download: Performance Report - ProDx Protease column [PDF]
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Product Data Sheet - ProDx Protease columns
Download: Product Data Sheet - ProDx Protease columns [PDF]
Also see: Part Number List - ProDx columns
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ProDx Protease columns
ProDx consumables are specifically designed and developed for use with the LEAP HDX platform.
The ProDx Protease column is optimized to perform at 0°C, the ideal temperature for performing HDX reactions.
Improved protease coupling reduces carryover from non-specific binding, while lower backpressures minimize the potential for contamination and overpressure leakage.
When used in combination with the new LEAP HDX valve system, it can be backflushed automatically to remove frit-loaded or undigested intact material. Clean columns give consistent retention times, overcoming traditional issues with non-selective binding and poor reproducibility.
To ensure confidence in your data, combine the ProDx Protease and trap columns for your HDX work
Features and benefits
- Improve workflow efficiencies and reduce instrument downtime
- Low temperature operation optimized at 0°C
- Lower carryover and increased desalting efficiency
- High pressure hardware designed for continuous use at 20,000 psi (1,379 bar)
- More overlapping fragments for high confidence data
- Automated backflushing when used with the LEAP HDX system
Recommended applications
Automated Hydrogen-Deuterium Exchange (HDX) experiments using the LEAP HDX platform.
Product specifications
ProDx columns were developed specifically for use with the LEAP HDX system. Packed in bio-inert stainless steel glass lined columns to withstand high pressure digestion practices, making it compatible with other systems.
- ID configuration: 2.1 mm
- Length configuration: 30 mm
The ProDx Protease (Pepsin) column is the first in a range of immobilized Protease columns available from Trajan. The initial intended use is with the LEAP HDX workflow, where these columns are used in-line for protein-protein and protein-ligand studies. The columns can be deployed outside of the HDX workflow as appropriate, possible alternate configurations might include general proteomics workflows, protein purification studies and QC applications. Column chemistry is such that limited exposure to common denaturing reagents and quenching salts (such as 6.4M Guanidine Hydrochloride, 8M Urea and 4M TCEP) do not adversely impact proteolytic performance.
ProDx Protease (Pepsin) columns are delivered chilled for optimal performance. Columns need to be maintained between 2-8°C. Column performance can be reduced if the column is maintained and operated above 15°C for extended periods of time, or if exposed to non-acidified aqueous reagents or organic reagents.
| Phase | Pore size (Å) | Particle size (µm) |
| Protease (Pepsin)* | 500-10,000 Å | 20 μm |
More
Please refer to the part number list for more items.
*This product contains POROS® material used under license for Sample Prep purposes from Life Technologies Corporation, a subsidiary of Thermo Fisher Scientific, Inc. The POROS® Trademark is owned by Thermo Fisher Scientific, Inc.
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ProDx trap columns
ProDx consumables are specifically designed and developed for use with the LEAP HDX platform.
ProDx trap columns are optimized for maximum recovery of peptide fragments, efficient desalting and low backpressures during the digestion and elution processes of HDX runs.
While many columns used for HDX trapping are not designed for use with low temperature HDX systems, ProDx trap columns are designed to operate down to 0°C, with consistent recovery, and demonstrate reduced backpressure at standard conditions of 200 μL/min, 100% Aqueous and 0°C, lowering instances of leakage and need for troubleshooting.
ProDx trap columns are pre-configured with PEEKsil® lines for ease of use, reducing the number of fittings needed in the system, preventing issues that commonly occur when relying on other columns with multiple connections.
To ensure confidence in your data, combine the ProDx Protease and trap columns for your HDX work
Features and benefits
- High recovery of peptide fragments generated by inline digestion
- Does not interfere with analytical separation
- Low temperature operation optimized at 0°C
- Lower carryover and increased desalting efficiency
- PEEKsil lines for reduced backpressure and leakage
- End frits designed to prevent rapid blocking by intact proteins and aggregates
- Phase and pore size to suit your analysis
Recommended applications
Automated Hydrogen-Deuterium Exchange (HDX) experiments using the LEAP HDX platform.
Product specifications
ProDx columns were developed specifically for use with the LEAP HDX system. ProDx trap columns are designed to protect the ProDx analytical column.
- ID configuration: 0.5 mm
- Length configuration: 20 mm
Currently available in C8 Analog*, with performance similar to C18 materials, suitable for reversed-phase separation of biomolecules such as proteins and peptides.
Fittings for ProDx trap columns
Choice of fittings available with ProDx trap columns, for low to ultra high pressure applications.
ProteCol® PEEK Fingertight fittings are precision manufactured to complement your HPLC needs.
| ProteCol® PEEK Fingertight HPLC fittings | Pressure rating | Maximum operating pressure rating when used with ProDx trap columns |
| 1/16" to 10-32 UNF (long) | 5,000 psi (345 bar) | 5,000 psi (345 bar) |
| 1/32" to 6-40 UNF | 2,750 psi (190 bar) | 2,750 psi (190 bar) |
PEEKLok™ UHPLC fittings are designed specifically for PEEKsil tubing: reliable, affordable, and easy to use, holding up to 22,000 psi; or up to 15,000 psi when used with ProDx trap columns.
| PEEKLok™ UHPLC fittings | Pressure rating | Maximum operating pressure rating when used with ProDx trap columns |
| 1/16" to 10-32 UNF | 22,000 psi (>1,500 bar) | 15,000 psi (1,000 bar) |
| 1/32" to 6-40 UNF | 22,000 psi (>1,500 bar) | 15,000 psi (1,000 bar) |
More
Please refer to the part number list for more items.
*This product contains POROS® material used under license for Sample Prep purposes from Life Technologies Corporation, a subsidiary of Thermo Fisher Scientific, Inc. The POROS® Trademark is owned by Thermo Fisher Scientific, Inc.
View full product details
Technical Poster - Automated Hydrogen-Deuterium Exchange with optimized consumables
HDX
Download: Technical Poster - Automated Hydrogen-Deuterium Exchange with optimized consumables [PDF]
Technical Poster - Trajan’s LEAP HDX platform is the leading automation solution for HDX-MS applications