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. 1968 Jan;14(1):69–79.

Antibody production studied by means of the localized haemolysis in gel (LHG) assay

II. Assay procedure

H H Wortis, R B Taylor, D W Dresser
PMCID: PMC1409262  PMID: 5635754

Abstract

The localized haemolysis in gel (LHG or plaque) assay has been investigated and it has been shown that the best condition for the assay is a plate made up from a standard tissue culture medium (199) in an agarose gel. Several convenient but sub-optimal alternatives are described. Other variables that were investigated were the concentration of indicator erythrocytes in the assay plate, the amount of complement added, the incubation time and the neutralization of xenogeneic developing serum by free mouse serum γ-globulins. The reasons for using log(x+1) transformed data for statistical analysis are also presented.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ARMITAGE P. Studies in the variability of pock counts. J Hyg (Lond) 1957 Dec;55(4):564–581. doi: 10.1017/s0022172400037451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. ASHWOOD-SMITH M. J. LOW TEMPERATURE PRESERVATION OF MOUSE LYMPHOCYTES WITH DIMETHYL SULFOXIDE. Blood. 1964 Apr;23:494–501. [PubMed] [Google Scholar]
  3. Cunningham A. J. A method of increased sensitivity for detecting single antibody-forming cells. Nature. 1965 Sep 4;207(5001):1106–1107. doi: 10.1038/2071106a0. [DOI] [PubMed] [Google Scholar]
  4. Dresser D. W., Wortis D. H. Use of an antiglobulin serum to detect cells producing antibody with low haemolytic efficiency. Nature. 1965 Nov 27;208(5013):859–861. doi: 10.1038/208859a0. [DOI] [PubMed] [Google Scholar]
  5. HALLIDAY W. J., WEBB M. A PLAQUE TECHNIQUE FOR COUNTING CELLS WHICH PRODUCE ANTIBACTERIAL ANTIBODY. Aust J Exp Biol Med Sci. 1965 Apr;43:163–166. doi: 10.1038/icb.1965.13. [DOI] [PubMed] [Google Scholar]
  6. Hege J. S., Cole L. J. A mathematical model relating circulating antibody and antibody forming cells. J Immunol. 1966 Jul;97(1):34–40. [PubMed] [Google Scholar]
  7. INGRAHAM J. S., BUSSARD A. APPLICATION OF A LOCALIZED HEMOLYSIN REACTION FOR SPECIFIC DETECTION OF INDIVIDUAL ANTIBODY-FORMING CELLS. J Exp Med. 1964 Apr 1;119:667–684. doi: 10.1084/jem.119.4.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Landy M., Sanderson R. P., Jackson A. L. Humoral and cellular aspects of the immune response to the somatic antigen of Salmonella enteritidis. J Exp Med. 1965 Sep 1;122(3):483–504. doi: 10.1084/jem.122.3.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Merchant B., Hraba T. Lymphoid cells producing antibody against simple haptens: detection and enumeration. Science. 1966 Jun 3;152(3727):1378–1379. doi: 10.1126/science.152.3727.1378. [DOI] [PubMed] [Google Scholar]
  10. Möller G. 19S antibody production against soluble lipopolysaccharide antigens by individual lymphoid cells in vitro. Nature. 1965 Sep 11;207(5002):1166–1168. doi: 10.1038/2071166a0. [DOI] [PubMed] [Google Scholar]
  11. NUSSENZWEIG R. S., MERRYMAN C., BENACERRAF B. ELECTROPHORETIC SEPARATION AND PROPERTIES OF MOUSE ANTIHAPTEN ANTIBODIES INVOLVED IN PASSIVE CUTANEOUS ANAPHYLAXIS AND PASSIVE HEMOLYSIS. J Exp Med. 1964 Aug 1;120:315–328. doi: 10.1084/jem.120.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Weiler E., Melletz E. W., Breuninger-Peck E. Facilitation of immune hemolysis by an interaction between red cell-sensitizing antibody and gamma-globulin allotype antibody. Proc Natl Acad Sci U S A. 1965 Nov;54(5):1310–1317. doi: 10.1073/pnas.54.5.1310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wortis H. H., Taylor R. B., Dresser D. W. Antibody production studied by means of the LHG assay. I. The splenic response of CBA mice to sheep erythrocytes. Immunology. 1966 Dec;11(6):603–616. [PMC free article] [PubMed] [Google Scholar]

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