A simple spectrophotometric method for estimating glycosaminoglycan concentrations
โ Scribed by Edeard W. Gold
- Publisher
- Elsevier Science
- Year
- 1979
- Tongue
- English
- Weight
- 381 KB
- Volume
- 99
- Category
- Article
- ISSN
- 0003-2697
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โฆ Synopsis
A rapid and simple spectrophotometric method is described for the estimation of microgram quantities of glycosaminoglycans following the formation of soluble complexes with alcian blue dye. The method is based on the different absorption spectra of the dye and dye-glycosaminoglycan complexes. No heating, centrifugation, lengthy equilibration, or sophisticated instrumentation which hamper other methods are required. Samples are mixed with freshly prepared dye solution and absorbance readings at 480 nm are compared to an appropriate standard curve. Albumin and individual monosaccharides do not interfere with the assay but high concentrations of chloride ion do. The method is suitable for the estimation of total glycosaminoglycan levels in biological fluids such as urine and blood.
Methods
Reagents. Alcian blue 8GN was purchased from Matheson, Coleman and Bell,
๐ SIMILAR VOLUMES
Human y-globulin, hemoglobin 2 x crystallized, purified globin, d-chymotrypsin 3~ crystallized, Nutritional Biochemicals Company; ribonuclease, crystallized, chromatographically pure, Mann Research Labs; lysosyme, 3 x crystallized, Sigma Chemical Co.
An ordinary spectrophotometer was used to study growth rates and increases in population size of nematodes by optical density measurements of nematodes suspended in 30-40% sucrose (w/v) solutions. The sucrose solution retarded the movement of nematodes in suspension and thereby decreased the fluctua