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Related Experiment Videos

Hemolytic plate assay for quantification of active human complement component C3 using methylamine-treated plasma as

M Ploug, T E Jessen, K G Welinder

    Analytical Biochemistry
    |May 1, 1985
    PubMed
    Summary
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    A new hemolytic plate assay accurately measures active human complement component C3 (complement component 3). This method is ideal for tracking C3 during purification and screening plasma samples for C3 levels.

    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • The human complement system is crucial for innate immunity.
    • Accurate quantification of complement components, like C3, is vital for diagnosing immune disorders.

    Purpose of the Study:

    • To develop and validate a specific hemolytic plate assay for active human complement component C3.
    • To establish a reliable method for C3 quantification in biological samples.

    Main Methods:

    • Utilized a hemolytic assay based on alternative pathway complement activation by rabbit erythrocytes.
    • Employed methylamine-treated plasma to provide complement components excluding C3.
    • Incubated at 37°C for 5 hours, with results stable upon overnight incubation.

    Main Results:

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  • The assay demonstrated linearity between lysis diameter and logarithm of C3 concentration (0.1-10x normal plasma levels).
  • Achieved a standard deviation below 10%, indicating high precision.
  • Hemolytic agarose plates are stable for 2 weeks at 4°C.

    • Conclusions:

    • The described hemolytic plate assay is specific and sensitive for active human C3.
    • The assay is suitable for C3 purification monitoring and plasma sample screening.
    • This method offers a cost-effective and convenient approach for C3 analysis.