产品名称：HYALURONIDASE, S. HYALURONLYTICUS NOV
Application Notes: Assay for Enzymatic Activity
• Prepare acetate buffer: 150 mM NaCl, 20 mM sodium acetate-acetic acid buffer, pH 6.0.
• Prepare hyaluronic acid solution: 50 mg purified potassium hyaluronate (containing approximately 38% glucuronic acid) is dissolved in 100 ml of the above mentioned sodium acetate buffer.
• Measure acidic bovine serum albumin with a spectrophotometer at 280 nm. Adjust the total absorbance of the albumin solution to 910. Dilute to 100 ml with 0.5 N acetate buffer, pH 4.3. Adjust the pH of the albumin solution to 3.1 using 4 N HCl. Boil in a water bath for 30 min at 100°C. Adjust to 100 ml with distilled H2O. This solution is stable for 3-4 weeks at 4°C. To use this solution, dilute with 5 volumes of acetate buffer from above.
• For the assay, mix 0.5 ml of the enzyme solution (made with acetate buffer without NaCl) with 0.5 ml of the hyaluronic acid solution. Incubate for 30 min at 60°C.
• Add 4 ml of the acidic bovine serum albumin solution. Incubate for 10 min at 60°C.
• Measure the turbidity at 660 nm. Obtain a blank value by using deactivated enzyme (incubate at 100°C for 10 min). Correct the reading by subtracting the blank value.
(blank value - measured value/blank value) x 2 x 2 x dilution = TRU/ml
Molecular Weight: 91000
Presentation: White lyophilized powder.
Contaminants: Protease: ≤ 0.05 units/ampule (pH 6.0)
Specific Activity: ≥2000 U/A280
Properties: Solubility: 500 mM Acetate buffer, pH 6.0 (30 U/ml) or H2O (30 U/ml).
Storage Conditions: Shipped at Ambient Temperature. Store at +2 - 8°C.