Quantitation of immobilized proteins

• Published in: Journal of biochemical and biophysical methods Vol. 21; no. 2; p. 129
• Main Authors: Lewis, W S, Schuster, S M
• Format: Journal Article
• Language: English
• Published: Netherlands 01-07-1990
• Subjects: Acrylic Resins; Collodion; Enzymes, Immobilized - analysis; Gels; Proteins - analysis; Rosaniline Dyes; Sepharose; Serum Albumin, Bovine - analysis
• ISSN: 0165-022X
• DOI: 10.1016/0165-022X(90)90060-P

A method for the quantitative determination of immobilized proteins based on the binding and subsequent elution of Coomassie Blue R is presented. Also presented is a method for the immobilization of proteins in solution by entrapment in polyacrylamide. These entrapped proteins are then available for use in the assay method presented. Other analytical procedures can also be performed on the entrapped proteins, either alone or in combination with the protein quantitation. The dye binding and elution method presented provides a sensitive and, in most applications, rapid method for the quantitative detection of immobilized proteins. Rather than immobilization being an obstacle to the assay method, this approach utilizes the advantages of immobilization for the removal of excess reagents. Application of this approach to several types of immobilized protein are presented.