Detection and quantitation of proteoglycans extracted from cell culture medium and cultured cartilage slices

Journal Article · Tue Nov 01 04:00:00 EST 1988 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(88)90050-4· OSTI ID:5990738

Hronowski, L J; Anastassiades, T P

Detection and quantitation of extracted proteoglycans, by staining with the dye Alcian blue on cellulose acetate followed by dissolution of the stained cellulose acetate strips in dimethyl sulfoxide containing 0.5% (v/v) sulfuric acid for absorbance measurement, is described. It is shown that, in the present system, the dye uptake by the proteoglycan is dependent only on the glycosaminoglycan content of the proteoglycan. The method is applied to the quantitation and characterization of proteoglycans and glycosaminoglycans, which have been extracted from radiolabeled bovine ankle cartilage and from mononuclear cell supernatant and which have been separated by DEAE-Sephacel column chromatography. The high sensitivity of the method allows detection of proteoglycans in 25-microliters samples of solutions containing as little as 1 microgram of glycosaminoglycan per milliliter of solution.

• Research Organization: Queen's Univ., Kingston, Ontario (Canada)
• OSTI ID: 5990738
• Journal Information: Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 174:2; ISSN ANBCA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CARTILAGE
CATTLE
CELL CULTURES
CHROMATOGRAPHY
CONNECTIVE TISSUE
DOMESTIC ANIMALS
DYES
GLYCOPROTEINS
ISOTOPE APPLICATIONS
ISOTOPES
LEUKOCYTES
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MATERIALS
MEASURING METHODS
ORGANIC COMPOUNDS
PROTEINS
RUMINANTS
SEPARATION PROCESSES
STAINS
SULFUR ISOTOPES
TISSUES
TRACER TECHNIQUES
VERTEBRATES