Specific Activity: ≥15 units/mg protein
Storage: 2-8°C
UNSPSC Code: 12352200
RIDADR: NONH for all modes of transport
Features and Benefits:
Free of impurities that may interfere with separation of peptides in RP HPLC.
Application:
Use Endoproteinase Glu-C Sequencing Grade for protein structure analysis and for sequence analysis.
General description:
Endoproteinase Glu-C Sequencing Grade is a serine protease isolated from Staphylococcus aureus V8 as a highly purified and specific protease. Inhibitors of Endoproteinase Glu-C are DFP, α2-macroglobulin and TLCK.
Other Notes:
For life science research only. Not for use in diagnostic procedures.
Preparation Note:
Working concentration: 1/100 to 1/20 (w/w)
With a minimum amount of 1 to 5 μg insulin Box, sufficient levels can be detected using the Column Aquapore RUP 300, 4.6 x 100 nm, 7 μm.
Working solution: Solvent is recommended in double-distilled water.
Solvents other than water have not been tested; Roche recommends using 50 mM Hepes, pH 7.8.
Storage conditions (working solution): -15 to -25 °C
At least 2 months if stored frozen in portions.
Stability: At +2 to +8°C within specification range for 12 months; stored dry.
Reconstitution:
Lyophilized Endoproteinase Glu-C sequencing grade is reconstituted in double-dist. water. To avoid autolysis, the incubation temperature should not exceed 25 °C.
Specificity:
Serine protease that very specifically hydrolyzes peptide bonds at the carboxylic side of Glu (in ammonium bicarbonate, pH 7.8, or ammonium acetate buffer, pH 4.0), or Glu and Asp (in phosphate buffer, pH 7.8). The specificity of Endoproteinase Glu-C Sequencing Grade is verified with the oxidized B-chain of insulin (insulin Box) as substrate. High concentrations of Endoproteinase Glu-C Sequencing Grade (1 part by weight enzyme with 10 parts by weight insulin Box) are incubated to detect traces of impurities, for example, contaminating proteases.
Heat inactivation: Most proteases sequencing grade are stopped by boiling for 5 minutes.