Immuno slot-blot assay using a membrane which covalently binds protein.
The use of a recently available commercial membrane (Immunodyne Immunoaffinity), which covalently binds proteins, in the development of a quantitative immunological slot blot assay is demonstrated. This membrane was found to be valuable in the quantification of polygalacturonase from tomato fruit, a protein which binds weakly to nitrocellulose membrane. The covalent binding of protein to a solid support allows the use of stringent conditions necessary for reduction of non-specific interactions between antibodies and other proteins. This method should prove useful to develop quantitative immunoassays for proteins which elute from nitrocellulose or other membranes during assay conditions.
Marlow
S J
SJ
Handa
A K
AK
D000906:Antibodies / Q000276:immunology; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D002021:Buffers; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005109:Extracellular Matrix; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D010940:Plant Proteins / Q000032:analysis*; D011096:Polygalacturonase / Q000032:analysis / Q000276:immunology; D011485:Protein Binding
D000906:Antibodies; D002021:Buffers; D010940:Plant Proteins; D003101:Collodion; D011096:Polygalacturonase