Endoproteinase GluC

Catalog # Concentration Size List Price Quantity Your Price
P8100S 50 µg $154.00
$138.60
Catalog # Size List Price Your Price
P8100S 50 µg $154.00
$138.60
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Endoproteinase GluC is a serine proteinase which preferentially cleaves peptide bonds C-terminal to glutamic acid residues. It also cleaves at aspartic acid residues at a rate 100-300 times slower than at glutamic acid residues.

  • Ideal for proteome analysis
  • Recombinant enzyme with no detectable non-specific protease contaminating activities
  • Free of glycerol and detergents which may interfere with Mass Spectrometry or liquid chromatography methods
  • Supplied in dry format
  • Optimal activity and stability for up to 12 months

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).

MALDI-TOF Ms
A 16 hour GluC digestion of Equine heart cytochrome c using a 1:20 enzyme to substrate ratio, followed by LC-MS with a Thermo™ LTQ Orbitrap XL, identified 97 peptides with 77% sequence coverage.
Product Source
Staphylococcus aureus Protease V8 gene cloned and expressed with histidine-tag in Bacillus subtilis.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Endoproteinase GluC P8100SVIAL -20 1 x 50 µg
  GluC Reaction Buffer B8100SVIAL -20 1 x 1.5 ml 2 X

Properties & Usage

Reaction Conditions

1X GluC Reaction Buffer
Incubate at 37°C

1X GluC Reaction Buffer
50 mM Tris-HCl
0.5 mM Glu-Glu
(pH 8 @ 25°C)

Usage Concentration

100 ng/μl

Molecular Weight

Theoretical: 29849 daltons


Notes
  • Storage conditions: Supplied in dry format from Tris-HCl and sodium chloride buffer.
  • Repeated freeze/thawing is not recommended.
  • 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.
  • 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.
References
  • 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.
  • Benner, J.S., Martin, D. and Schampel, A.K. unpublished results.
  • 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.
  • 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.
Selection Charts
Quality Control Assay
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Specifications
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Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
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