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Hyaluronan Isolation from Mouse Mammary Gland.

Bio-protocol (2018-06-05)
Cornelia Tolg, Mary Cowman, Eva A Turley
ABSTRACT

The glycosaminoglycan hyaluronan (HA) is a key component of the extracellular matrix. The molecular weight of HA is heterogeneous and can reach from several million to several hundred daltons. The effect of HA on cell behavior is size dependent; fragmented HA acts as a danger signal, stimulates cell migration and proliferation and is proinflammatory, native high molecular weight HA suppresses inflammation. Therefore, it is important to analyze HA size distribution when studying the role of HA in tissue homeostasis and pathology. This protocol describes isolation of HA from mouse mammary glands but can also be applied to other tissues. The quality of the isolated HA is sufficient to analyze size distribution by gel electrophoresis ( Calabro et al., 2000 ).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Calcium chloride dihydrate, for molecular biology, ≥99.0%
Millipore
Benzonase® Nuclease, ≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution
USP
Deferoxamine mesylate, United States Pharmacopeia (USP) Reference Standard