Journal article
Detection and enumeration of immunoglobulin secreting cells
Journal of immunological methods, Vol.124(1), pp.35-42
1989
DOI: 10.1016/0022-1759(89)90183-X
PMID: 2478637
Abstract
A method is described in which sheep red blood cells (SRBC) are coated with anti-immunoglobulin (Ig) antibodies (Ab) for use in reverse hemolytic plaque assays (RHPA) as follows. The non-complement fixing F(ab′)
2 fragments of rabbit anti-mouse Ig Ab are derivatized with the
N-hydroxysuccinimide ester of palmitate. The hydrophobic palmitate tails spontaneously insert into the SRBC membranes, thus coating the cells with anti-Ig F(ab′)
2 molecules. The SRBC are lysed by successive additions of mouse Ig, rabbit anti-mouse Ig and complement. When this procedure is carried out in agar gel, Ig-secreting mouse cells produce localized hemolytic areas (plaques). The procedure is more reproducible and more sensitive than RHPA performed with protein A-coated SRBC. In principle, this procedure should be adaptable to the detection of cells secreting any molecule for which specific antibodies are available.
Details
- Title: Subtitle
- Detection and enumeration of immunoglobulin secreting cells
- Creators
- Annette J SchlueterMariangela SegreDiego Segre
- Resource Type
- Journal article
- Publication Details
- Journal of immunological methods, Vol.124(1), pp.35-42
- Publisher
- Elsevier B.V
- DOI
- 10.1016/0022-1759(89)90183-X
- PMID
- 2478637
- ISSN
- 0022-1759
- eISSN
- 1872-7905
- Language
- English
- Date published
- 1989
- Academic Unit
- Pathology
- Record Identifier
- 9984046901302771
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