产品信息

• 产品组分信息

  本产品提供以下试剂或组分：

  NEB #	名称	组分货号	储存温度	数量	浓度
  P8113S     -20       α-Lytic Protease P8113SVIAL -20 1 x 20 µg 0.4 mg/ml
  P8113L     -20       α-Lytic Protease P8113SVIAL -20 5 x 20 µg 0.4 mg/ml

• 特性和用法

  贮存溶液

  10 mM sodium acetate
  pH 5 @ 25°C

  热失活

  95°C

  分子量

  实际: 19.8 kDa

• 优势和特性

  Features

  • Digestion of proteins for proteomic analysis by mass spectrometry
  • Protein and peptide identification
  • Site-specific identification and quantitation of endogenous SUMO modifications under native conditions⁽¹⁾. 
  • Cleavage after aliphatic amino acids resulting in increased sequence coverage of membrane proteins 

• 相关产品

  相关产品

  • Endoproteinase GluC
  • Trypsin-ultra™, Mass Spectrometry Grade
  • Endoproteinase AspN
  • Endoproteinase LysC

• 注意事项

  1. α-Lytic Protease is stable for at least 2 years at –20°C. No loss of activity is observed after 10 freeze-thaw cycles.
  2. To avoid autolysis, α-Lytic Protease should be stored long term in 10 mM Sodium Acetate pH 5.0. 
  3. 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.
  4. Enzyme activity is stimulated in up to 0.1% sodium deoxycholate⁽²⁾. 
  5. 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⁾.
  6. 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⁾.
  7. 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.

• 参考文献

  1. Lumpkin, RJ. et al. (2017). Nat Commun.. 8 (1), 1171.
  2. Meyer, JG. et al. (2014). Molecular & Cellular Proteomics. 13 (3), 823-835.
  3. Ruse, C. New England Biolabs, Inc.. Unpublished observation

操作说明、说明书 & 用法

• 操作说明

  1. α-Lytic Protease In-gel Digestion Protocol (NEB #P8113)
  2. α-Lytic Protease Typical Reaction Protocol (NEB #P8113)

工具 & 资源

• 选择指南

  • Protease Selection Chart

FAQs & 问题解决指南

• FAQs

  1. What reaction buffers are recommended for use with α-Lytic Protease?
  2. Can I perform my digest with α-Lytic Protease at temperatures other than 37°C?
  3. Why should I use α-Lytic Protease instead of Trypsin?
  4. Is there an advantage to treating my sample with both α-Lytic Protease and Trypsin?
  5. What reagents inhibit α-Lytic Protease activity?
  6. What reagents stimulate α-Lytic Protease activity?
  7. Can α-Lytic Protease be heat-inactivated?