English
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.
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.
Other Notes
One unit will degrade 0.75 μg of the polysaccharide hyaluronic acid per minute at pH 5.35 at 37 ºC (as measured by turbidimetric absorbance (λ: 600nm) when complexed with BSA after 45 minutes).
