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Native hyaluronidase from Streptomyces hyaluronlyticus nov. species. Catalyzes the degradation of hyaluronic acid. Produces δ4,5-unsaturated tetra- and hexasaccharides.
Catalogue Number
389561
Brand Family
Calbiochem®
References
References
Baker, J.R., et al. 2002. Biochem. J.365, 317. Mogelerskii, M.S. 1964. Fed. Proc. 23, T559.
Product Information
CAS number
37326-33-3
Activity
≥ 100 U/ampule [1 U = 1 TRU (Turbidity Reducing Unit)]
Unit of Definition
One TRU (Turbidity Reducing Unit = 1 U) is defined as the amount of enzyme that will hydrolyze 50% of hyaluronic acid, determined by a 50% reduction in turbidity, in 30 min at 60°C pH 6.0.
Following reconstitution, store in the refrigerator (4°C). Stock solutions prepared in water are stable for up to 2 weeks at 4°C and stock solutions prepared in 500 mM acetate buffer, pH 6.0, are stable for up to 1 month at 4°C.
Baker, J.R., et al. 2002. Biochem. J.365, 317. Mogelerskii, M.S. 1964. Fed. Proc. 23, T559.
数据表
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
07-July-2009 JSW
Description
Native hyaluronidase from Streptomyces hyaluronlyticus nov. species. Catalyzes the degradation of hyaluronic acid. Produces δ4,5-unsaturated tetra- and hexasaccharides.
Please Note: The appearance of this product can vary from gray cottony fibers to balled up droplets with no change in quality.
Form
White lyophilized powder
Recommended reaction conditions
Assay for Enzymatic ActivityReagent Preparation• 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.
Activity Protocol• 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
CAS number
37326-33-3
EC number
3.2.1.35
Contaminants
Protease: ≤ 0.05 units/ampule (pH 6.0)
Specific activity
≥2000 U/A280
Activity
≥ 100 U/ampule [1 U = 1 TRU (Turbidity Reducing Unit)]
Unit definition
One TRU (Turbidity Reducing Unit = 1 U) is defined as the amount of enzyme that will hydrolyze 50% of hyaluronic acid, determined by a 50% reduction in turbidity, in 30 min at 60°C pH 6.0.
Solubility
500 mM Acetate buffer, pH 6.0 (30 U/ml) or H₂O (30 U/ml)
Storage
+2°C to +8°C
Do Not Freeze
Ok to freeze
Special Instructions
Following reconstitution, store in the refrigerator (4°C). Stock solutions prepared in water are stable for up to 2 weeks at 4°C and stock solutions prepared in 500 mM acetate buffer, pH 6.0, are stable for up to 1 month at 4°C.
Toxicity
Standard Handling
Merck USA index
14, 4758
References
Baker, J.R., et al. 2002. Biochem. J.365, 317. Mogelerskii, M.S. 1964. Fed. Proc. 23, T559.
