A cautionary note on the use of lentil lectin affinity chromatography in the presence of sodium deoxycholate.
When affinity chromatography with lentil lectin (LcH)-Sepharose was carried out in 0.5% (w/v) sodium deoxycholate (DOC), 10 mm Tris, pH 8.2, LcH consistently appeared in the 0.1 M alpha-methyl-mannoside (alpha-MeMan)-eluted fraction. Eluted LcH was recovered in both the control and specific immunoprecipitates with antisera directed against components of the glycoprotein fraction, suggesting that at least a portion of the LcH retained the capacity to interact with protein-bound carbohydrate. We suggest that the presence of functional LcH in the purified glycoprotein fraction is a potential source of artifact which may not be generally appreciated. When the non-ionic detergent. Nonidet P-40 was substituted for DOC in the same type of LcH-Sepharose fractionation, LcH did not appear in the 0.1 M alpha-MeMan-eluted fraction.
D000818:Animals; D000941:Antigens; D001345:Autoradiography; D002458:Cell Fractionation; D002846:Chromatography, Affinity / Q000379:methods*; D003840:Deoxycholic Acid / Q000494:pharmacology*; D006023:Glycoproteins / Q000276:immunology; D006041:Goats; D037102:Lectins / Q000494:pharmacology*; D008198:Lymph Nodes / Q000166:cytology; D037121:Plant Lectins; D013552:Swine
D000941:Antigens; D006023:Glycoproteins; D037102:Lectins; D037121:Plant Lectins; C033619:lentil lectin; D003840:Deoxycholic Acid