Glycoproteins and antigens of membranes prepared from rat thymocytes after lysis by shearing or with the detergent Tween-40.
In purification of cell surface antigens an efficient method for preparing membrane from large numbers of cells is needed. Such a method is described for preparing membranes from rat thymocytes after lysis in the non-ionic detergent Tween-40. Cell surface antigens were recovered at a yield of 30-50%, and a purification of 30-40-fold. By contrast enzyme markers for the other cell organelles were present in the membrane fraction in very low yield. The membrane obtained with the detergent method was compared with that resulting from the best of previously describes methods involving cell lysis by shearing. The detergent method compared favourably for simplicity as well as for yield and purification, and both membrane preparations contained similar protein and glycoprotein constituents. The main glycoprotein bands of membranes from thymocytes and thoracic duct lymphocytes were identified after polyacrylamide gel electrophoresis in dodecyl sulphate. In thymocyte membrane, three main bands at apparent molecular weights of 150 000, 84 000 and 25 000 were seen, and of these the 84 000 glycoprotein did not bind to the lentil lectin. In thoracic duct lymphocyte membrane the 25 000 glycoprotein was absent and a band at 95 000 was intensified in comparison with thymocytes.
D000818:Animals; D000941:Antigens / Q000032:analysis*; D002462:Cell Membrane / Q000032:analysis* / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis*; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000032:analysis*; D008970:Molecular Weight; D010743:Phospholipids / Q000032:analysis; D011136:Polysorbates; D051381:Rats; D013601:T-Lymphocytes / Q000032:analysis* / Q000276:immunology; D013950:Thymus Gland / Q000032:analysis* / Q000276:immunology
D000941:Antigens; D006023:Glycoproteins; D008565:Membrane Proteins; D010743:Phospholipids; D011136:Polysorbates