Description(s):
Heat shock protein 60 (HSP60) is a family of heat shock proteins that prevent proteins from misfolding during stressful situations such as high heat. HSP60 functions as a molecular chaperone, which either helps improperly folded proteins refold into their correct conformation or tags them for degradation. HSP60 levels are typically low during normal physiological conditions and increase in response to cellular stress.
Principle
The wells of the plate are coated with capture antibody specific to HSP60. In this assay, the test sample initially reacts with the solid phase capture antibody, resulting in HSP60 being bound to the well. The wells are then washed to remove unbound proteins, and biotin-linked antibodies are added to bind to the immobilized HSP60. After washing away the unbound antibodies, Streptavidin-HRP conjugate is added to form a complex with the antibody-bound HSP60. After incubation, the wells are washed to remove unbound Streptavidin-HRP conjugate. TMB substrate is then added and forms a blue color when the HRP-linked antibodies are detected. The reaction is then terminated with Stop Solution, which changes the color from blue to yellow. The HSP60 concentration in each well is directly proportional to its color intensity and can be quantified by measuring its optical density at 450 nm (OD450) in a microplate reader.
