The serum complement system is a series of dissolved proteins that can defend against a variety of pathogens. There are three independent but related pathways for complement activity: the classical pathway, the alternative pathway and the lectin pathway. Each complement pathway is a protease cascade. These three pathways have different initial pathways, but share the downstream effector functions necessary for inflammation and pathogen clearance. The alternative pathway and the lectin complement pathway function independently without adaptive immunity. In contrast to these other pathways, the classical complement pathway is activated in the presence of specific antibodies. The activity of complement in serum can be determined by its ability to lyse red blood cells in vitro.
In this assay, sheep red blood cells are used as indicator cells. The red blood cells (RBC; also called red blood cells) of vertebrates contain large amounts of the cytoplasmic respiratory pigment hemoglobin, which is easily detected after being released from the cells. The lysis of red blood cells turns the experimental diluent red, and the intensity of the red color (equal to the amount of hemoglobin released) can be measured photometrically. Activation of complement to cleavage SRBC results in the release of hemoglobin into the buffer. The target SRBC that has not been lysed by complement is separated from the supernatant by centrifugation, and the intensity of red in the supernatant is quantified by measuring the absorbance of the solution at 540 nm. The measured absorbance is related to the amount of hemoglobin released due to complement-mediated hemolysis (disruption of red blood cells).
Turnaround time is ~ 4 weeks
1 ml frozen serum (see testing requisition for specimen handling).
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Creative Biolabs provides luciferase-based ADCC assay. This Jurkat cell based assay is pioneered by Creative Biolabs, and the methodology is very well accepted by the field. See attached ADCC Reporter Assay Protocol for further details.
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