Fluorescence linked immunosorbant assays using microtiter plates.
Fluorescence methods are widely used in the detection of antibodies and other binding events. However, as a general screening and detection tool in microtiter plates, enzyme linked immunosorbant (ELISA) methods predominate. In this paper we explore all parameters for effective use of fluorescence as a plate based detection method, including which microtiter plates can be used, the most effective means of immobilization, and the use of different fluorescent dyes or fluorescent proteins. These studies indicate that fluorescent immunosorbant assays (FLISA) can be used as effectively as enzymatic method in microtiter plate based screening methods, including the screening of phage antibody selections.
B division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, United States.
N
N
Velappan
J
J
Clements
C
C
Kiss
R
R
Valero-Aracama
P
P
Pavlik
A R M
AR
Bradbury
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001360:Avidin / Q000276:immunology; D005456:Fluorescent Dyes; D015200:Fluoroimmunoassay / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007163:Immunosorbent Techniques; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; C093483:neutravidin; D001360:Avidin; D049452:Green Fluorescent Proteins