A novel ELISA using PVDF microplates.
Here we describe the development of a novel specific, rapid ELISA system, which is performed on modified microplates where polyvinylidine fluoride (PVDF) forms the base of each well. The use of microplates with PVDF membranes as the solid phase allows for a greater binding capacity of protein in comparison to the solid phases of traditional ELISAs. The increased binding capacity of the solid phase provides for the direct binding of antigens, which can subsequently be assayed using a single, specific and well-characterized antibody. This direct assay system eliminates the need for two distinct antibodies that are often necessary in conventional two site ELISA systems. The system is able to specifically detect purified proteins as well as antigens in crude preparations of tissue homogenates. The PVDF-based ELISA performs with similar sensitivity and reproducibility as conventional two site ELISAs in tissue homogenates. The intra- and inter-assay coefficients of variation for the measurement of actin in crude rat brain homogenate were 2.36 and 5.15%, respectively.
Halim
Nader D
ND
Clinical Brain Disorders Branch, Intramural Research Program, National Institute of Mental Health, NIH, Bethesda, MD 20892-1385, USA.
Joseph
Andrew W
AW
Lipska
Barbara K
BK
D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism*; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008422:Materials Testing; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D018482:Muscle, Skeletal / Q000378:metabolism*; D011145:Polyvinyls / Q000737:chemistry*; D011817:Rabbits; D051381:Rats; D014018:Tissue Distribution; D005465:Fluorocarbon Polymers
D000199:Actins; D020099:Coated Materials, Biocompatible; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers