Literature DB >> 6423576

An automatic fluorescence micro-ELISA system for quantitative screening of hybridoma supernatants using a protein-A-beta-galactosidase conjugate.

P L Van Soest, J de Josselin de Jong, P M Lansdorp, W Van Ewijk.   

Abstract

We report on a rapid micro-ELISA screening procedure for the detection of monoclonal antibodies directed against cell surface determinants on lymphoid cells of the mouse. This method employs Terasaki-type trays coated with a monolayer of target cells fixed with 0.02% glutaraldehyde. Cells are incubated with monoclonal antibodies, followed by affinity column-purified rabbit-anti-rat immunoglobulin antibodies and a protein-A-beta-galactosidase conjugate. Binding of antibodies to the cells is visualized by incubation with the substrate 4-methylumbelliferyl galactoside. Fluorescence in the individual wells of the Terasaki trays is then quantitatively analysed within 120 s using a scanning inverted microfluorometer, connected to a digital voltmeter and a desk-top calculator.

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Year:  1984        PMID： 6423576     DOI： 10.1007/bf01003433

Source DB:  PubMed          Journal:  Histochem J        ISSN： 0018-2214


  27 in total

1.  Expression of MHC antigens by mouse thymic dendritic cells.

Authors:  R V Rouse; W van Ewijk; P P Jones; I L Weissman
Journal:  J Immunol       Date:  1979-06       Impact factor: 5.422

2.  Monoclonal antibodies in transplantation research.

Authors:  A F Williams
Journal:  Transplantation       Date:  1979-03       Impact factor: 4.939

3.  Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters.

Authors:  B Rotman; B W Papermaster
Journal:  Proc Natl Acad Sci U S A       Date:  1966-01       Impact factor: 11.205

4.  Detection of HLA antigens on blood platelets and lymphocytes by means of monoclonal antibodies in an ELISA technique.

Authors:  P M Lansdorp; F Oosterhof; G C Astaldi; W P Zeijlemaker
Journal:  Tissue Antigens       Date:  1982-01

5.  A comparative study on the preparation of immunoglobulin-galactosidase conjugates.

Authors:  A M Deelder; R de Water
Journal:  J Histochem Cytochem       Date:  1981-11       Impact factor: 2.479

6.  Use of an enzyme-linked immunosorbent assay (ELISA) for screening of hybridoma antibodies against cell surface antigens.

Authors:  L Suter; J Brüggen; C Sorg
Journal:  J Immunol Methods       Date:  1980       Impact factor: 2.303

7.  Distribution of H-2 microenvironments in the mouse thymus. Immunoelectron microscopic identification of I-A and H-2K bearing cells.

Authors:  W Van Ewijk; R V Rouse; I L Weissman
Journal:  J Histochem Cytochem       Date:  1980-10       Impact factor: 2.479

8.  The use of quantiatative cytochemical analyses in rapid prenatal detection and somatic cell genetic studies of metabolic diseases.

Authors:  H Galjaard; A Hoogeveen; W Keijzer; E De Wit-Verbeek; C Vlek-Noot
Journal:  Histochem J       Date:  1974-09

9.  Immunoelectron microscopy of cell surface antigens: a quantitative analysis of antibody binding after different fixation protocols.

Authors:  W Van Ewijk; R C Coffman; I L Weissman
Journal:  Histochem J       Date:  1980-05

10.  Automated measurement of immunogalactosidase reactions with a fluorogenic substrate by the aperture defined microvolume measurement method and its potential application to Schistosoma mansoni immunodiagnosis.

Authors:  A M Deelder; G Koper; R De Water; H J Tanke; J P Rotmans; J S Ploem
Journal:  J Immunol Methods       Date:  1980       Impact factor: 2.303
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  2 in total

1.  Quantitation of dye binding by cell monolayers in a microtiter system.

Authors:  M R Barer; H Lyon; B S Drasar
Journal:  Histochem J       Date:  1986 Feb-Mar

2.  Loss of antibody binding to prefixed cells: fixation parameters for immunocytochemistry.

Authors:  W Van Ewijk; P L Van Soest; A Verkerk; J F Jongkind
Journal:  Histochem J       Date:  1984-02
  2 in total

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