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H4272

Sigma-Aldrich

Hyaluronidase from bovine testes

Type IV-S, powder, suitable for mouse embryo cell culture, 750-3000 units/mg solid

Synonym(s):
Hyaluronate 4-glycanohydrolase, Hyaluronoglucosaminidase
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.75

sterility

aseptically processed

Quality Level

type

Type IV-S

form

powder

specific activity

750-3000 units/mg solid

mol wt

~55 kDa (four subunits of 14 kDa each)

composition

Protein, 80-105% biuret

technique(s)

cell culture | embryo: suitable

storage temp.

−20°C

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Application

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

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).

Preparation Note

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.

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificate of Analysis

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Certificate of Origin

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