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Quantitation of Component Molecules of Precipitated Class II Complexes

  • J. Lindsay
  • J. F. Maddox
  • J. G. Bodmer

Abstract

The use of a scanning instrument that detects β emissions from dried gels has enabled us to quantitate emission levels from individual class II products. If radiolabeled molecules are presented as discrete spots or bands on an SDS/PAGE gel, they can be detected and analyzed. The instrument used is an AMBIS Mark II β scanner (Automated Microbiology Systems, Inc.), which detects β particles passed through a mylar filter into an argon-methane mix. Localization of each ionization event is determined, and a two-dimensional representation is built up. The scanner is operated from an IBM PC AT-X, which allows data to be stored and retrieved for subsequent analysis. We examined profiles from cells expressing class II products to determine whether the location of the epitope on the dimer had an effect on the ratio of α and β chains bound by monoclonal antibody. We then used this to examine the expression of α and β chains in P815-transfected cells.

Keywords

Lymphoblastoid Cell Line Component Molecule Discrete Spot Free Pool Imperial Cancer Research Fund 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Ozato K, Mayer N, Sachs DH. Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and la antigens. J Immunol 1980; 124: 533–540.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1989

Authors and Affiliations

  • J. Lindsay
    • 1
  • J. F. Maddox
    • 1
  • J. G. Bodmer
    • 1
  1. 1.Imperial Cancer Research FundLondonUK

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