Lysyl EndopeptidaseR, Mass Spectrometry Grade (Lys-C)

Grade :	for Proteome Research
Storage Condition :	Keep at -20 degrees C.

Manufacturer : FUJIFILM Wako Pure Chemical Corporation

CAS RN® : 123175-82-6

Catalogue/Product Number	Package Size	COA Download
125-05061	20ug x5
121-05063	20ug

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SDS

Product Specification Sheet

Overview

Outline	This product is for research use only. Do not administer it to human. [BIOCHEMISTRY] For Proteome Research! Among the most important techniques in proteome analyses is the in-gel digestion of protein spots/bands that have been resolved by electrophoresis using digestive enzymes, such as trypsin and lysyl endopoptidase. Proteins can be identified by mass spectrometry analysis of the peptides produced by in-gel digestion, and further information regarding post-translational modifications can be obtained.Lysyl Endopeptidase, Mass Spectrometry Grade is a freeze dried product that retained sufficient activity for in-gel digestion and packed in very small quantities for convenience purposes. [Features] High specificity and efficiency of protein digestion allow for easy database searches by peptide mass. Improved cleavage at lysine residue and increase in the number of peptides are obtained by combination with trypsin. Packed in very small quantities according to the amounts used so that sufficient activity for in-gel digestion may be retained. [Related Products] Wako Catalog #293-57701 Negative Gel Stain MS Kit, for Electrophoresis (20 tests) Wako Catalog #299-58901 Silver Gel Stain MS Kit, for Electrophoresis (20 tests) [Physical properties] Appearance: Lyophilized form containing ca. 10% Tris-HCl buffer, pH 8. Activity: Shown on each label Molecular Weight: 27,000 (gel filtration); 30,000 (SDS electrophoresis) Solubility: Soluble in water or buffer solution. Stability: Stable at 4 degrees C, when dissolved in buffer of pH 5~12. Stable at 30 degrees C in the range of pH 6~11, but unstable at 50 degrees C or higher. Optimal pH: 9.0 -9.5 (Amidase activity) Isoelectric point: 6.9~7.0 Substrate specificity: Inhibitors: DFP, PMSF, TLCK [References] Wada, Y., and Kadoya, M.: J. Mass Spectrom., 38, 117 (2003). Shevchenko, A., Wilm, MM., Vorm, O., and Mann., M.: Anal. Chem., 68, 850 (1996). Source: Bacteria Originally, the source of this product was indicated as "Achromobacter lyticus" based on the physiological and morphological properties of the bacteria. However, we confirmed that the 16SrDNA sequence was highly homologous to that of Lysobacter

Outline

This product is for research use only. Do not administer it to human.

[BIOCHEMISTRY]

For Proteome Research!

Among the most important techniques in proteome analyses is the in-gel digestion of protein spots/bands that have been resolved by electrophoresis using digestive enzymes, such as trypsin and lysyl endopoptidase. Proteins can be identified by mass spectrometry analysis of the peptides produced by in-gel digestion, and further information regarding post-translational modifications can be obtained.Lysyl Endopeptidase, Mass Spectrometry Grade is a freeze dried product that retained sufficient activity for in-gel digestion and packed in very small quantities for convenience purposes.

[Features]

High specificity and efficiency of protein digestion allow for easy database searches by peptide mass.
Improved cleavage at lysine residue and increase in the number of peptides are obtained by combination with trypsin.
Packed in very small quantities according to the amounts used so that sufficient activity for in-gel digestion may be retained.

[Related Products]

Wako Catalog #293-57701 Negative Gel Stain MS Kit, for Electrophoresis (20 tests)
Wako Catalog #299-58901 Silver Gel Stain MS Kit, for Electrophoresis (20 tests)

[Physical properties]

Appearance: Lyophilized form containing ca. 10% Tris-HCl buffer, pH 8.
Activity: Shown on each label
Molecular Weight: 27,000 (gel filtration); 30,000 (SDS electrophoresis)
Solubility: Soluble in water or buffer solution.
Stability: Stable at 4 degrees C, when dissolved in buffer of pH 5~12. Stable at 30 degrees C in the range of pH 6~11, but unstable at 50 degrees C or higher.
Optimal pH: 9.0 -9.5 (Amidase activity)
Isoelectric point: 6.9~7.0
Substrate specificity:
Inhibitors: DFP, PMSF, TLCK

[References]

Wada, Y., and Kadoya, M.: J. Mass Spectrom., 38, 117 (2003).
Shevchenko, A., Wilm, MM., Vorm, O., and Mann., M.: Anal. Chem., 68, 850 (1996).

Source: Bacteria

Originally, the source of this product was indicated as "Achromobacter lyticus" based on the physiological and morphological properties of the bacteria. However, we confirmed that the 16SrDNA sequence was highly homologous to that of Lysobacter

Application

Comparison of In-gel Digestion using Trypsin (Tp), Lysl Endopeptidase (Lep) and Lep Combined with Tp (Lep + Tp)

BSA band (100 ng) resolved by SDS-PAGE was in-gel digested with Tp, Lep and Lep + Tp and analyzed by MALDI-TOFMAS. The figure shows the individual mass spectra. The evaluation of these peptidases is summarized in the table.

Mass spectrum using Tp and Tp + Lep

When Tp is used concurrently with Lep, additional peaks were found around m/z 2000 compared to when only Tp is used. It shows that the recovery rate of peptide increases.

Table. Comparison ofTp, Lep, and Tp + Lep

	Tp	Lep	Tp + Lep
Cleavage site	C-terminal of Arg and Lys	C-terminal of Lys	C-terminal of Arg and Lys
Missed Cleavage(Rates of missed cleavage)※	Many (8 %)	Very few (0 %)	Few (3 %)
Number of identified peptides	17	19	22

※The coverage (percentage of available peptides in the entire sequence) obtained by database search using a parameter setting of Missed Cleavage = 0 was subtracted from the coverage obtained by using a parameter setting of Missed Cleavage = 1.The value resulted from subtracting the coverage obtained when database searches were performed with Missed cleavage 0 from that obtained when performed with Missed cleavage 1. “Coverage” is the percentage of peptides obtained after in-gel digestion in the whole sequence.

These results indicate there are very few missed cleavages obtained by Lep digestion. When Tp is used concurrently with Lep, missed cleavages decrease and the number of identified peptides increase compared to when only Tp is used.

Data wasprovided by Dr. Y. Wada, Osaka Medical Center and Research Institute for Maternal and Child Health.

Property

Appearance	Lyophilisate
Origin / Source	Bacteria
Activity	0.03 - 0.07AU/vial

Alias

Lys-C