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A method for efficient and selective recovery of membrane glycoproteins from concanavalin A-Sepharose using media containing sodium dodecyl sulfate and urea

✍ Scribed by Laurent Poliquin; Gordon C. Shore


Publisher
Elsevier Science
Year
1980
Tongue
English
Weight
564 KB
Volume
109
Category
Article
ISSN
0003-2697

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✦ Synopsis


Herpes-specific membrane glycoproteins were recovered from infected cells by incubating total homogenates with Con A-Sepharose in sealed plastic tubes. Following affinity binding of glycoproteins, subsequent washes with media containing high-salt concentrations followed by washes in 0.1% sodium dodecyl sulfate effectively removed nonglycoprotein contaminants. Glycoproteins were then eluted in high yield by heating the Con A-Sepharose-glycoprotein complex in medium containing 5% sodium dodecyl sulfate and 8 M urea. Eluates were placed directly onto sodium dodecyl sulfate-polyacrylamide gels for further analysis and purification of individual components. The procedure described here is convenient for simultaneously processing many different samples on either a large or small scale.