产品信息

Endoproteinase GluC (Staphylococcus aureus Protease V8) is a serine proteinase which selectively cleaves peptide bonds C-terminal to glutamic acid residues (1). Endoproteinase GluC also cleaves at aspartic acid residues at a rate 100-300 times slower than at glutamic acid residues (2,3).

产品来源

Staphylococcus aureus Protease V8 gene cloned and expressed with histidine-tag in Bacillus subtilis

重组

Endoproteinase GluC should be reconstituted by the addition of 50-500 µl of high purity water. Finger flick the volume of water in the tube to fully resuspend the enzyme. Rapid autolysis is a function of enzyme concentration; any sample reconstituted in a small volume should be used immediately. To get the most use out the enzyme, resuspend the enzyme in 500 μl H₂O and aliquot 50 ul each in 10 tubes. Freeze the tubes that are not being used immediately at -20°C for up to two weeks or less. Storage at – 80°C will prolong enzyme stability approximately 2-4 additional weeks.

产品类别:

Proteases Products,

Proteome Analysis Products

应用:

Glycomics and glycoproteomics,

Protein Digestion

• 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
  P8100S     -20       Endoproteinase GluC P8100SVIAL -20 1 x 50 µg Not Applicable   GluC Reaction Buffer B8100SVIAL -20 1 x 1.5 ml 2 X

• 特性和用法

  反应条件

  1X GluC Reaction Buffer
  Incubate at 37°C

  2X GluC Reaction Buffer
  50 mM Tris-HCl
  0.5 mM Glu-Glu
  (pH 8 @ 25°C)

  使用浓度

  100 ng/μl

  分子量

  理论上的: 29849 daltons

• 相关产品

  相关产品

  • Trypsin-ultra™, Mass Spectrometry Grade
  • Endoproteinase AspN
  • Endoproteinase LysC
  • α-Lytic Protease

• 注意事项

  1. Storage conditions: Supplied in dry format from Tris-HCl and sodium chloride buffer.
  2. Repeated freeze/thawing is not recommended.
  3. GluC may also be used in 20-50mM phosphate or ammonium bicarbonate buffer. For optimal activity, supplement reactions with Glu-Glu dipeptide. Glutamic acid residues are highly favored in Tris, phosphate and ammonium bicarbonate buffers.
  4. GluC is purified with a His-tag at its C-terminal of the protein. The predominant species has four histidine residues at the C-terminus with the remaining species showing a distribution from one to six His tags.

• 参考文献

  1. Drapeau, G.R., Boily, Y. and Houmard, J. (1972). Purification and properties of an extracellular protease of Staphylococcus aureus. J. Biol. Chem.. 247, 6720-26.
  2. Benner, J.S., Martin, D. and Schampel, A.K. unpublished results.
  3. Birktoft, J.J. and Breddam, K. (1994). Proteolytic Enzymes: Serine and Cysteine Peptidases. A.J. Barrett(Ed.), Glutamyl endopeptidases. Methods Enzymol.. 244, 114-126. San Diego: Academic Press.
  4. Breddam, K. and Meldal, M. (1992). Substrate preference of glutamic-acid-specific endopeptidases assessed by synthetic peptide substrates based on intramolecular fluorescence quenching. Eur. J. Biochem.. 206, 103-7.

操作说明、说明书 & 用法

• 操作说明

  1. Typical GluC Digest Reaction Conditions (P8100)
  2. Typical Trypsin and GluC Co-Digest Reaction Conditions (P8100)
  3. Typical GluC In-Gel Digest Reaction Conditions (P8100)

工具 & 资源

• 选择指南

  • Protease Selection Chart

FAQs & 问题解决指南

• FAQs

  1. What is the stability/compatibility of Endoproteinase GluC with regards to presence of DTT, urea, concentrations of methanol, acetonitrile, nitrosoguanidine, thiourea and glycerol?
  2. How does one carry out limited proteolysis experiments with Trypsin and Endoproteinase Glu-C? Which chemicals can be used to quench the reactions at given time points to monitor proteolysis progress via SDS- PAGE?