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Related Experiment Videos

An improved sensitive and simple microassay of mouse complement.

R M De Waal1, G Schrijver, M J Bogman

  • 1Department of Pathology, University Hospital Nijmegen, The Netherlands.

Journal of Immunological Methods
|April 6, 1988
PubMed
Summary
This summary is machine-generated.

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A new microassay quickly measures mouse complement activity using sheep red blood cells. Protein A purification of antibodies ensures accurate classical pathway complement determination in serum samples.

Area of Science:

  • Immunology
  • Biochemistry
  • Assay Development

Background:

  • Classical pathway complement activity is crucial for immune responses.
  • Accurate measurement of complement activity in mouse serum is essential for research.
  • Existing methods can be time-consuming or require large sample volumes.

Purpose of the Study:

  • To develop a simple, fast, and sensitive hemolytic microassay for classical pathway complement activity in mouse serum.
  • To optimize antibody preparations for reliable complement-mediated hemolysis.
  • To enable high-throughput determination of CH50 titers.

Main Methods:

  • Hemolytic microassay using antibody-sensitized sheep red blood cells (SRBC).
  • Photometric measurement of hemolysis at 405 nm using an ELISA plate scanner.

Related Experiment Videos

  • Protein A purification of anti-SRBC IgG to remove anticomplementary factors and enrich IgG2 antibodies.

    • Main Results:

    • A simple and rapid hemolytic microassay was successfully developed.
    • Protein A purification of antibodies yielded optimal and standardized results.
    • The assay demonstrated high sensitivity for determining CH50 titers in small mouse serum samples (80 µL).
    • The method allows for a large number of tests to be performed in a single day.

    Conclusions:

    • The developed microassay provides a sensitive and efficient method for assessing classical pathway complement activity in mouse serum.
    • Optimized antibody preparation is critical for accurate and reproducible complement assays.
    • This rapid, one-step assay facilitates high-throughput screening of complement function in immunological studies.