By J.G. Beeley
The previous 20 years have noticeable a spread of curiosity in glycoproteins. From being a borderline region among carbohydrate and protein chemistry, it has turn into correct to a variety of organic phenomena.
The goal of the ebook is to explain thoughts which might be used to respond to the various uncomplicated questions on glycosylated proteins. equipment are mentioned for isolation; compositional research; constitution carbohydrate devices; protein-carbohydrate linkages, conserving in brain the various nature of difficulties which readers can have to tackle.
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Additional resources for Glycoprotein and Proteoglycan Techniques
In some cases it may be possible to isolate glycoprotein from a single type of cell grown in culture. The quantity of material obtainable from cell culture will necessarily be limited but radioactive labelling techniques (Chapter 8) can be employed to increase the sensitivity of detection. Genetic variation in glycoproteins can be minimised by examination of glycoprotein from a single (preferably homozygous) individual. 3. Extraction and solubilisation Many glycoproteins, such as those which occur in serum, are freely soluble in dilute salt solutions and will remain in solution unless the protein moiety is denatured.
5 M methyl a-D-glucopyranoside in deoxycholate buffer. In this preparation only part of the Thy-1 antigen binds to lentil lectin. The activity which is not bound can be purified by antibody affinity chromatography. 5. 01 M NaCl(4 ml/g tissue) and homogenised in a Waring blender for 2 min.
Effects of shearing forces and sonication Prolonged Ch. , 1966). However, material extracted in this way is degraded. Sadjera and Hascall (1969) showed that the average sedimentation coefficient of proteoglycan was decreased by the shearing forces during high-speed homogenisation. , 1967). Homogenisation under milder conditions may nevertheless be valuable in promoting the solubilisation of proteoglycan (Brandt and Muir, 1971). Sonication may, in addition to shearing forces, give rise to free radicals which could potentially degrade protein or carbohydrate moieties of proteoglycans.
Glycoprotein and Proteoglycan Techniques by J.G. Beeley