Immunoblot analysis of the eighth component of human complement. Demonstration of subunits and detection of C8α-γ double and triple bands

W. Nürnberger, R. Seger, P. Kobler, I. Mons, F. Tedesco, V. Wahn

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Using SDS-PAGE/immunoblot analysis of the eighth component of human complement, C8, we have been able to demonstrate an 85 kDa C8α-γ and a 62 kDa C8β subunit in normal human serum. Serum from an undiagnosed patient who presented undetectable hemolytic C8 activity possessed only the 85 kDa subunit, suggesting a defect in the C8β subunit. Serum of a patient with known C8α-γ deficiency possessed only the complementary 62 kDa subunit. Both sera used together were able to lyse antibody-sensitized sheep erythrocytes, whereas individual sera could not. Optimum conditions for C8 immunoblotting were determined using small amounts of serum or plasma, during low voltage electrophoresis and a sensitive staining technique (nitrobluetetrazolium/bromochloroindoxylphosphate). Using these conditions, the C8α-γ subunit was found to be composed of up to three bands, termed C8α-γ1, -2 and -3. All three bands were found in pooled normal sera. Individual sera had at least the C8α-γ2 and C8α-γ3 bands. Two C8β-deficient sera from two unrelated patients exhibited only the C8α-γ2 and C8α-γ3 bands. We conclude that immunoblotting of C8 permits a detailed analysis of the molecular composition of this component and helps to establih a precise diagnosis in inherited C8 deficiencies.

Original languageEnglish
Pages (from-to)257-263
Number of pages7
JournalJournal of Immunological Methods
Volume109
Issue number2
DOIs
Publication statusPublished - May 9 1988

Fingerprint

Serum
Immunoblotting
Complement C8
Electrophoresis
Polyacrylamide Gel Electrophoresis
Sheep
Erythrocytes
Staining and Labeling
Antibodies

Keywords

  • C8 deficiency
  • C8α-γ variant
  • Inherited complement deficiency
  • Neisserial infection

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Immunoblot analysis of the eighth component of human complement. Demonstration of subunits and detection of C8α-γ double and triple bands. / Nürnberger, W.; Seger, R.; Kobler, P.; Mons, I.; Tedesco, F.; Wahn, V.

In: Journal of Immunological Methods, Vol. 109, No. 2, 09.05.1988, p. 257-263.

Research output: Contribution to journalArticle

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