Fixation of cell-bound antibody in the membrane immunofluorescence test

Biberfeld, Peter; Biberfeld, Gunnel; Molnar, Zelma; Fagræus, Astrid

doi:10.1016/0022-1759(74)90056-8

Cited by 44 publications

(15 citation statements)

References 15 publications

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“…The improved sensitivity is primarily due to the use of better equipment, particularly the microscope, light source, and objectives, and also to the very advantageous use of paraformaldehyde-Triton-fixed instead of acetonefixed cell cultures. Paraformaldehyde-Triton has been reported to be the least denaturating reagent for fixation of cell cultures for fluorescence microscopy (Biberfeld et al 1974). Results demonstrating that IF was more sensitive than 50°/~PNT have already been reported (Olesen & Jsrgensen 1984).…”

Section: Discussionmentioning

confidence: 92%

Detection of rainbow trout antibody to Egtved virus by enzyme-linked immunosorbent assay (ELISA), immunofluorescence (IF), and plaque neutralization tests (50%PNT)

Olesen¹,

Lorenzen²,

Jørgensen³

1991

Dis. Aquat. Org.

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ABSTRACT. The humoral immune response to Egtved virus, the causative agent of viral haemorrhagic septicaemia (VHS) in rainbow trout Oncorhynchus mykiss, was investigated by means of the enzymelinked immunosorbent assay (ELISA), immunofluorescence (IF), a n d the 50 'X plaque neutralization test (50°/"PNT). Sera from fish immunized with virus under aquarium conditions as well a s sera collected from fish in farms with different VHS status were included In the experiments. ELISA proved to b e more sensitive and less time-and material-consuming than either IF or 50°/~PNT Using Elisa, antibody to Egtved virus was found in 54 %I of fish from infected trout farms examined w h~l e only 16 % were positive by 50°/r,PNT The IF test had a sensitivity close to that of ELISA, but was less suitable for examination of large numbers of sera. Non-neutralizing antibodies tend to perslst longer after VHS infection than neutralizing antibodies. The kinetics of the antibody response was close to that prev~ously published. No anamnestic immune response was observed with any of the tests. We conclude that our ELISA is well suited for VHS surveillance.

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Section: Discussionmentioning

confidence: 92%

Detection of rainbow trout antibody to Egtved virus by enzyme-linked immunosorbent assay (ELISA), immunofluorescence (IF), and plaque neutralization tests (50%PNT)

Olesen¹,

Lorenzen²,

Jørgensen³

1991

Dis. Aquat. Org.

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“…Paraformaldehyde, unlike acetone, leaves the cell membrane impermeable to antibodies (Biberfeld et al, 1974) and thus enables antibody-mediated detection of viral proteins only on the surface of infected cells. In cell cultures fixed with paraformaldehyde each of the three G-specific MAbs stained plaques consisting of 20 to 50 infected cells (Table 2), whereas MAbs to the other structural proteins (N, M 1 and M2) of Egtved virus were unable to stain at all, except for a few disrupted cells (not shown).…”

Section: Effect Of Tunicamycin On Immunofluorescenee Staining Patternsmentioning

confidence: 99%

Neutralization of Egtved virus pathogenicity to cell cultures and fish by monoclonal antibodies to the viral G protein

Lorenzen¹,

Olesen²,

Jørgensen³

1990

Journal of General Virology

122

113

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“…[2] with monova lent anti-IgE. Apparently, the fixation with PFA prevents redistribution of IgE after in cubation with the antiserum [3], while after incubation of nonfixed cells at 0 °C still some redistribution occurs, although this was not found by Sundqvist [19]. This pos sibility could explain the contrast between the forementioned linear configuration and the patchy distribution found by Sullivan el al.…”

Section: Discussionmentioning

confidence: 90%

Electron Microscopic Studies on Human Basophils from Atopic and Nonatopic Subjects, Using Horse Radish Peroxidase Labelled Anti-IgE

Elven¹,

Stallman²,

Brühl³

1977

Int Arch Allergy Immunol

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In previous studies, quantitative differences in cell-bound IgE on the basophils from various donors were measured by means of a quantitative immunofluorescence technique. In this study, cell-bound IgE was demonstrated on basophilic granulocytes by immunoelectron microscopy, using horse radish peroxidase labelled anti-IgE. The distribution of IgE on the basophils was studied in both atopic and nonatopic subjects. The intensity of the peroxidase staining on the basophils varied and appeared to correlate with the amount of cell-bound IgE, as estimated by quantitative immunofluorescence.

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Fixation of cell-bound antibody in the membrane immunofluorescence test

Cited by 44 publications

References 15 publications

Detection of rainbow trout antibody to Egtved virus by enzyme-linked immunosorbent assay (ELISA), immunofluorescence (IF), and plaque neutralization tests (50%PNT)

Detection of rainbow trout antibody to Egtved virus by enzyme-linked immunosorbent assay (ELISA), immunofluorescence (IF), and plaque neutralization tests (50%PNT)

Neutralization of Egtved virus pathogenicity to cell cultures and fish by monoclonal antibodies to the viral G protein

Electron Microscopic Studies on Human Basophils from Atopic and Nonatopic Subjects, Using Horse Radish Peroxidase Labelled Anti-IgE

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