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H4272 Sigma-Aldrich

Hyaluronidase from bovine testes

Type IV-S, powder, mouse embryo tested, 750-3000 units/mg solid

Synonym: Hyaluronate 4-glycanohydrolase, Hyaluronoglucosaminidase

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Properties

Related Categories 3.2.x.x Glycosidases, 3.x.x.x Hydrolases, Application Index, Biochemicals and Reagents, Carbohydrate Hydrolysis,
sterility   aseptically processed
type   Type IV-S
form   powder
mol wt   ~55 kDa (four subunits of 14 kDa each)
composition   Protein, 80-105% biuret
suitability   mouse embryo tested
storage temp.   −20°C

Description

Application

Hyaluronidase is an enzyme derived from bovine testes. It is used to remove the cumulus mass from newly ovulated eggs, when preparing the eggs for in vitro culture.

Reconstitute 1 vial of hyaluronidase with 3 ml of M2 Medium (Product Nos. M 7167 or M 5910) or deionized water to obtain a 10 mg/ml stock solution. Dilute stock solution with M2 Medium to obtain a final working concentration of approximately 300 μg/ml.

Gonadotropins are used to superovulate 20 female mice. The mice are then mated and preimplantation stage embryos are retrieved and counted. The embryos are treated with hyaluronidase until the cumulus mass is dissociated from the embryos. The embryos are cultured, viewed daily and progress of the embryos is recorded.

Biochem/physiol Actions

Hyaluronidase degrades hyaluronan and has been found to be inappropriately regulated during cancer progression.

These enzymes randomly cleave β-N-acetylhexosamine-[1→4] glycosidic bonds in hyaluronic acid, chondroitin, and chondroitin sulfates.

Unit Definition

One unit will cause a change in % transmittance at 600 nm of 0.330 per minute at pH 5.35 at 37 °C in a 2.0 mL reaction mixture (45 minute assay).

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

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Certificate of Origin

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Cell Culture Guide
Protocols & Articles

Articles

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Protocols

Enzymatic Assay of Hyaluronidase (3.2.1.35)

This procedure is a turbidimetric determination (% Transmittance at 600 nm, Light path = 1 cm) of Hyaluronidase activity. It is based on the following reaction:
Keywords: Extinction coefficient

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Peer-Reviewed Papers
15

References

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