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Related Experiment Videos

Isolation and study of murine C3

M B Pepys, A C Dash, A H Fielder

    Immunology
    |October 1, 1977
    PubMed
    Summary

    Researchers successfully isolated purified mouse complement component 3 (C3) from serum and plasma. This native C3 demonstrated functional activity and underwent specific cleavage, yielding a distinct product in vitro.

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    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • Complement component 3 (C3) is a critical protein in the innate and adaptive immune systems.
    • Understanding C3 structure and function is vital for immune response research.

    Purpose of the Study:

    • To isolate purified, native mouse complement component 3 (C3) from serum and plasma.
    • To characterize the functional activity and cleavage products of isolated mouse C3.

    Main Methods:

    • Isolation of C3 via euglobulin precipitation and immunoadsorbent chromatography.
    • Functional assays including C3 fixation on EAC142gp cells and interaction with lymphocyte C3 receptors.
    • Molecular weight determination and analysis of cleavage products using SDS-PAGE and electrophoresis.

    Main Results:

    • Achieved 30-55% recovery of purified, native mouse C3.
    • Demonstrated functional activity of isolated C3 through cell-based assays.
    • Identified a major C3 cleavage product (≥210,000 MW) with altered electrophoretic mobility compared to in-plasma cleavage.

    Conclusions:

    • Established a reliable method for isolating functional, native mouse C3.
    • Characterized key properties of mouse C3, including its molecular weight and cleavage behavior.
    • Highlighted differences in C3 cleavage products depending on the context (isolated vs. plasma).

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