A rapid method for quantitation of immunofluorescence on human lymphoblastoid cell suspensions. |
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Authors: | T P Gillis F H Wilson L A Wilson J J Thompson |
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Institution: | Department of Microbiology, Louisiana State University Medical Center, 1100 Florida Avenue, New Orleans, Louisiana 70119 USA |
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Abstract: | A quantitative fluoroimmunoassay for antibodies to, and surface antigens of, human lymphoblastoid cells (IM-1) with photon-counting spectrofluorometry is described. IM-1 cell suspensions were reacted with rabbit antiserum to human spleen vesicular membranes, were washed, and then were reacted with an excess amount of fluorochrome-conjugated (fluorescein or rhodamine) goat anti-rabbit immunoglobulin G (IgG). Under appropriate conditions, antibodies to IM-1 cells could be detected with experimental/control fluorescence ratios ranging between 5 and 40. Moreover, detectable levels of antibody-saturated cells approached 5 × 103 cells per milliliter or a total of 1.7 × 103 cells per assay. Inhibition of the fluoroimmunoassay was performed with either viable IM-1 cells or IM-1 vesicular membrane preparations and demonstrated a dose-dependent antigen inhibition. Fluorescence of sensitized cells reactive with either fluorescein- or rhodamine-labeled antiglobulins could be quantitatively distinguished in dual-labeled preparations. |
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