The complement system consists of dozens of proteins that are present in different forms in the blood or bound to cells. These proteins are intimately linked to complement activation. The system is divided into three pathways of activation: the lectin pathway (LP), the classical pathway (CP) and the alternative pathway (AP), each with different recognition molecules. Here Creative Biolabs shares a brief introduction to the CP hemolytic assay protocol.
Classical Pathway of Hemolytic Assay Protocol
In CP assays, the time required for the maximum amount of C3 convertase to present on the cell surface is referred to as Tmax. Tmax is closely related to the amount of functionally active C4b on the cell surface and should usually be less than 6 minutes. Tmax was determined by incubating EAC14b cells with C2 and measuring the number of functionally active sites (Z) of the cells at specific times after the addition of C2.
Fig.1 Flow chart of determination of Tmax assay.
After Tmax was determined by the above assay, decay activity can be determined by the following procedure.
Fig.2 Assay of decay activity.
The diverse complement detection services provided by Creative Biolabs can deepen the understanding of the mechanism of disease development and promote the progress of complement-related drug development projects. Our professional team will provide customized solutions for your complement program according to your unique needs, and help your drug development progress in the early stage. In addition, rigorously tested complement products are also the best choice for your complement research.
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Please note that our protocols are only for your reference.
For Research Use Only.
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