Complement-Mediated Hemolysis Assay
Sheep red blood cells are employed as indicator cells in this test. The cytoplasmic respiratory pigment hemoglobin, which is readily detectable after it is released from the cells, is present in significant quantities in the red blood cells (RBCs; also known as red blood cells) of vertebrates. The experimental diluent turns red due to the lysis of red blood cells, and photometric measurement can be used to determine the intensity of the red color, which is a measure of the amount of hemoglobin released. Hemoglobin is released into the buffer upon activation of complement to cleave SRBC. Centrifugation is used to separate the target SRBC that has not been lysed by complement from the supernatant, and the absorbance of the solution at 540 nm is used to measure the amount of red in the supernatant. The quantity of hemoglobin produced as a result of complement-mediated hemolysis (disruption of red blood cells) is correlated with the observed absorbance.
