α-Lytic Protease (aLP) cleaves after Threonine (T), Alanine (A), Serine (S) and Valine (V) residues. Its specificity makes it an orthogonal and alternative protease to others commonly used in proteomics applications, including trypsin and chymotrypsin.
Analyze complex proteomes
Suitable for both in-gel and solution digests
Optimal activity and stability for up to 24 months
Ideal for digestion of proteins for proteomic analysis by mass spectrometry
Product Information
α-Lytic Protease (aLP) cleaves after Threonine (T), Alanine (A), Serine (S) and Valine (V) residues. Its specificity makes it an orthogonal and alternative protease to others commonly used in proteomics applications, including trypsin and chymotrypsin. Peptides generated by aLP are of similar average length to those of Trypsin.
Figure 1. α-Lytic Protease can be used alone or in combination with other proteases to yield increased sequence coverage.
Comparison of sequence coverage for three protein standards after parallel digestion using Trypsin (blue) and a-Lytic Protease (gold). The combined data set (grey) results in overlapping peptides and increased sequence coverage.
Product Source
Purified from Lysobacter enzymogenes
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α-Lytic Protease is stable for at least 2 years at –20°C. No loss of activity is observed after 10 freeze-thaw cycles.
To avoid autolysis, α-Lytic Protease should be stored long term in 10 mM Sodium Acetate pH 5.0.
α-Lytic Protease is active in a variety of mass spectrometry compatible buffers including ammonium bicarbonate, Tris-HCl, and HEPES. The optimal pH range for α-Lytic Protease digestion is pH 7.5 - 8.5.
Enzyme activity is stimulated in up to 0.1% sodium deoxycholate(2).
Enzyme activity is inhibited by 1.0% sodium deoxycholate (~60% active); 0.1% SDS (~50 active); 1% SDS (~40% active); 1 M guanidine hydrochloride (~20% active); 4 M guanidine hydrochloride (no activity); Serine protease inhibitors, such as PMSF (no activity)(2).
α-Lytic Protease is compatible with both in-solution and in-gel digestion methods. Digestion with α-Lytic Protease yields increased peptide recovery for standard and crushed gel conditions of large molecular weight proteins(3).
α-Lytic Protease is active over a wide temperature range from 4ºC – 50ºC, with optimal activity at 37ºC. Lower temperatures may require longer incubation times.
References
Lumpkin, RJ. et al. (2017). Nat Commun.. 8 (1), 1171.
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