产品名称：ENDOPEPTIDASE LYS-C, A. LYTICUS
Application Notes: Amidase Activity:
•Reagent A: 200 mM AMP, pH 9.5 (dissolve 4.2 g 2-amino-2-methyl-1,3-propanediol in 150 ml H2O; adjust the pH to 9.5 with 1 M HCl; add H2O to a final volume of 200 ml)
•Reagent B: 2.5 mM Bz-Lys-pNA (dissolve 22.6 mg Nα-benzoyl-DL-lysine-p-nitroanilide in 900 ml H2O)
•Reagent C: enzyme diluent (dissolve 240 µg 2-amino-2-hydroxymethyl-1,3-propanediol in 900 ml H2O; adjust the pH to 8.0 with 0.1 M HCl; add H2O to a final volume of 1 L
•Reagent D: enzyme solution (dissolve 1 vial enzyme in 1 ml H2O; dilute 100 µl to a final volume of 25 ml with Reagent C)
•45% Acetic Acid (stop solution)
1. Add 2.6 ml Reagent A to 300 µl Reagent B and pre-incubate at 30°C to equilibrate the temperature.
2. Add 100 µl Reagent D and incubate for 25 min at 30°C. Include 100 µl Reagent C alone as a control.
3. Add 1 ml 45% acetic acid to stop the reaction.
4. Measure the absorbance at 405 nm.
5. Calculate the activity:
Amidase Activity (AU/vial) = [(Atest-Acontrol)/25] x [1/9.62] x [4/0.1] x 250
Molecular Weight: 30000
Properties: Solubility: H2O or Tris-HCl buffer, pH 4-11. Reconstitute 1 vial with 1 ml solvent; if the enzyme is not completely solubilized, add 1-3 ml additional solvent until completely dissolved.
Specific Activity: ≥2 AU/mg protein
Storage Conditions: Shipped on Blue Ice. Store at -20°C. Avoid freeze/thaw.