Product
Tryp-N Sequencing kit
Tryp-N Sequencing enzyme
Size
10µg/20µg
10µg/20µg
Catalogue#
K-001
E-001
List Price
$250-$350
$225-$325
Tryp-N™
Tryp-N™ is a thermophilic metalloprotease with N-terminal specificity for arginine and lysine developed at Cold Spring Harbor Laboratory (CSHL). By placing basic centers of peptides (the amino terminus and side chains of K/R) in one location, b-ions predominate in MS/MS.
Benefits:
- Digestions can be completed in minutes to hours, not over night.
- Easy-to-read spectra. Tryp-N™ provides the complimentary b-ion series that can be missing from trypsin, allowing you to more easily find and specifically localize PTMs.
- Ion current is not diluted between termini (like with trypsin), enhancing sensitivity for example in MRM experiments (see below).
- Easily control digestion with the addition of EDTA for reproducible partial digests and increased sequence coverage.
- Tryp-N™ cleavage is unaffected by proline.
- It also doesn’t care about methylation.
- At higher temperatures (its Topt is 65 °C), proteins are denatured, assisting in the analysis of difficult-to-digest proteins.
- Tryp-N™ is highly stable across a wide variety of experimental conditions including pH (5-11), buffers, ionic strength and detergents (80% active in 0.5% SDS at 50 °C!).
- At higher temperatures, endogenous enzymes are denatured and inactive: useful if pesky phosphatases or proteases interfer with your analysis.
Left: digestion is complete in 0.5 – 4 hrs, depending on temperature. Middle: Ion current is exactly inverted between trypsin and Tryp-N™. Right: Limits of detection (LODs) in MRM experiments were determined for 17 peptides of catalase produced with trypsin or Tryp-N™. The median fold increase in sensitivity was 4.5x. Average = 35.4x. 50% CI = 1.96x – 18.6x increase.