Skip to main content
Immunology logoLink to Immunology
. 1969 Dec;17(6):999–1006.

Labelling of mouse alloantibody with tritiated DL-alanine

U Hämmerling, N Shigeno, L J Old, E A Boyse
PMCID: PMC1456028  PMID: 5386641

Abstract

Radio-iodination is not a satisfactory method of labelling mouse antibody, which is peculiarly susceptible to destruction during the process of iodination. An alternative which causes very little loss of antibody is the attachment of a tritium-labelled amino acid to mouse γG by peptide linkage. This is accomplished by reaction of γG with DL-alanine N-carboxy anhydride under mild conditions. The method is applicable to estimation in vitro of the relative amounts of H-2 antibody absorbed by viable cells, and of the relative amounts of H-2 antigen on viable cells. Non-specific uptake is virtually eliminated by: (i) prior absorption of the labelled product in vivo, (ii) pre-incubation of the cells in 20 per cent foetal bovine serum and its inclusion in the suspending medium, and (iii) performance of absorption procedures in the cold.

Full text

PDF
999

Selected References

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

  1. Boyse E. A., Stockert E., Old L. J. Isoantigens of the H-2 and Tla loci of the mouse: interactions affecting their representation on thymocytes. J Exp Med. 1968 Jul 1;128(1):85–95. doi: 10.1084/jem.128.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fuchs S., Sela M. Preparation and characterization of poly-DL-alanyl rabbit gamma-globulin. J Biol Chem. 1965 Sep;240(9):3558–3567. [PubMed] [Google Scholar]
  3. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  4. Humphrey J. H., Askonas B. A., Auzins I., Schechter I., Sela M. The localization of antigen in lymph nodes and its relation to specific antibody-producing cells. II. Comparison of iodine-125 and tritium labels. Immunology. 1967 Jul;13(1):71–86. [PMC free article] [PubMed] [Google Scholar]
  5. Kekwick R. A. The serum proteins in multiple myelomatosis. Biochem J. 1940 Sep;34(8-9):1248–1257. doi: 10.1042/bj0341248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. McFARLANE A. S. Efficient trace-labelling of proteins with iodine. Nature. 1958 Jul 5;182(4627):53–53. doi: 10.1038/182053a0. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES