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38594 Hyaluronidase, Ovine Testes

38594
  
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Replacement Information
Description
Overview

This product has been discontinued.



Native hyaluronidase from ovine testes. Hyaluronic acid degrading endoglycosidase that catalyzes the hydrolysis of 1,4 linkages in chondroitin, chondroitin-4-sulfate, chondroitin-6-sulfate esters, and hyaluronic acid.

Note: 1 KU = 1000 units.
Catalogue Number38594
Brand Family Calbiochem®
References
ReferencesLokeshwar, V.B., et al. 2002. J. Biol. Chem. 277, 33654.
Borders, C.L., and Raftery, M.A. 1968. J. Biol. Chem. 243, 3756.
DeSalequi, M., et al. 1967. Arch. Biochem. Biophys. 121, 548.
Lowry, O.H., et al. 1951. J. Biol. Chem. 193, 265.
Product Information
CAS number37326-33-3
Unit of DefinitionOne unit is defined as the amount of enzyme that will cause the same turbidity reduction as 1.0 unit of International Standard preparation.
EC number3.2.1.35
FormWhite lyophilized solid
Quality LevelMQ100
Applications
Biological Information
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Ambient Temperature Only
Toxicity Standard Handling
Storage -20°C
Do not freeze Ok to freeze
Special InstructionsFollowing reconstitution, store in the refrigerator (4°C). Stock solutions are stable for up to 5 days at 4°C.
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalogové číslo GTIN
38594 0

Documentation

Hyaluronidase, Ovine Testes MSDS

Title

Safety Data Sheet (SDS) 

Hyaluronidase, Ovine Testes Certificates of Analysis

TitleLot Number
38594

References

Přehled odkazů
Lokeshwar, V.B., et al. 2002. J. Biol. Chem. 277, 33654.
Borders, C.L., and Raftery, M.A. 1968. J. Biol. Chem. 243, 3756.
DeSalequi, M., et al. 1967. Arch. Biochem. Biophys. 121, 548.
Lowry, O.H., et al. 1951. J. Biol. Chem. 193, 265.
Data Sheet

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.

Revision26-Febuary-2009 RFH
DescriptionNative hyaluronidase from ovine testes. Catalyzes the hydrolysis of 1,4 linkages between N-acetylhexosamine and D-glucuronate residues in chondroitin, chondroitin-4-sulfate, chrondroitin-6-sulfate esters, and hyaluronic acid. Has an optimal pH of 4.5-6.0. Hydrolyzes the endo-N-acetyl-hexosaminic bonds of hyaluronic acid and chondroitin sulfate A and C to yield tetrasaccharide residues. Hyaluronidase activity is inhibited by the presence of Fe2+, Fe3+, Cu2+, and Mn2+.
FormWhite lyophilized solid
Recommended reaction conditions
Assay Protocol Principle of Assay: Hyaluronic acid is measured by its ability to yield turbidity with an acidic albumin solution. The turbidity produced is a function of hyaluronic acid concentration and hence can be related to the activity of the enzyme. Reagents: 1N NaOH 0.3 M Sodium Phosphate Buffer, pH 5.3-5.35 Hyaluronic Acid Solution (0.2 mg/ml in 0.3 M sodium phosphate buffer, pH 5.3-5.35). May heat in a boiling water bath for a few minutes to dissolve. Alternatively, place at 4°C overnight. Stir for about 1 h on the next day to dissolve completely. Enzyme Diluent (0.02 M sodium phosphate buffer, pH 6.9, containing 0.45% NaCl and 0.01% BSA) 5N Hydrochloric Acid Acid Albumin Solution (10 mg of BSA, Fraction V, in 10 ml of 0.5 M sodium acetate buffer, pH 4.2). Adjust pH to 3.75 with few drops of 50 mM HCl. This solution is stable for 2 days at 4°C. Standard Hyaluronidase Solution (10 I.U./ml). Enzyme solution: Dissolve 10 mg of Hyaluronidase in 2 ml of ice-cold enzyme diluent (see above). Dilute to 2.0 I.U./ml just prior to use. Test Procedure: (Incubation temperature: 38°C; Wavelength; 600 nm; d = 1 cm) Preparation of Standard curve: Into a series of test tubes add reagents, as follows:

Table 1: Preparation of Standard Curve

1. Thoroughly mix and place the tubes at 38°C for 5 min. 2. At time zero add 1 ml of hyaluronic acid solution to each tube (for convenience allow 30 s intervals between each tube). Mix, stopper and incubate at 38°C for exactly 45 min. 3. Then add 10 ml of acid albumin solution to each tube and mix rapidly by inversion. Allow to stand for 5 min at room temperature. Read absorbance at 600 nm against a blank. 4. Construct a standard curve by plotting absorbance against the I.U./ml. Measurement of Hyaluronidase Activity: Pipette 1 ml of each diluted sample into test tubes at 38°C and repeat steps 1 to 4 (see above). Obtain hyaluronidase concentration (I.U./ml) for each sample solution using the standard curve.

Figure 1: Calculations

Protein Determination: Protein Content can be determined by Lowry's method.

CAS number37326-33-3
EC number3.2.1.35
Unit definitionOne unit is defined as the amount of enzyme that will cause the same turbidity reduction as 1.0 unit of International Standard preparation.
SolubilityReconstitute at 5 mg/ml in 20 mM sodium phosphate, 0.45% NaCl, and 0.01% BSA, pH 6.9. At a concentration of 5 mg/ml the solution will be clear and colorless.
Storage -20°C
Do Not Freeze Ok to freeze
Special InstructionsFollowing reconstitution, store in the refrigerator (4°C). Stock solutions are stable for up to 5 days at 4°C.
Toxicity Standard Handling
Merck USA index14, 4758
ReferencesLokeshwar, V.B., et al. 2002. J. Biol. Chem. 277, 33654.
Borders, C.L., and Raftery, M.A. 1968. J. Biol. Chem. 243, 3756.
DeSalequi, M., et al. 1967. Arch. Biochem. Biophys. 121, 548.
Lowry, O.H., et al. 1951. J. Biol. Chem. 193, 265.