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Isolation of the fourth component (C4) of rat complement
Journal of Immunology (Baltimore, Md. : 1950)
|November 1, 1979
Summary
Researchers purified rat complement component 4 (C4) to homogeneity. They developed a monospecific antibody and determined normal plasma C4 concentration in Wistar rats.
Area of Science:
- Biochemistry
- Immunology
- Proteomics
Background:
- The complement system is crucial for innate and adaptive immunity.
- Rat complement component 4 (C4) plays a significant role in complement activation.
- Understanding C4 levels is vital for immunological research.
Purpose of the Study:
- To purify rat complement component 4 (C4) to homogeneity.
- To develop a monospecific antibody for rat C4.
- To quantify C4 concentration in normal rat plasma.
Main Methods:
- Sequential chromatography (QAE-A50, SP-C50, hydroxyapatite, gel filtration) of rat plasma.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for purity assessment.
- Antibody production in rabbits and characterization.
Main Results:
- Rat C4 was purified to homogeneity with a molecular weight of 198,000 Da.
- A monospecific antibody against rat C4 was successfully generated.
- Normal plasma concentration of rat C4 was determined as 190 ± 34 μg/ml.
Conclusions:
- Rat C4 can be effectively purified using a multi-step chromatographic approach.
- The developed antibody is suitable for detecting rat C4.
- Established normal C4 levels provide a baseline for future studies in rat models.