Human Heat Shock 60kD Protein 1, Chaperonin ELISA Kit

The Antibodies.com Human HSP60 ELISA Kit is a sandwich enzyme-linked immunosorbent assay (sELISA) intended for the in vitro quantitative determination of human Heat Shock 60kD Protein 1, Chaperonin (HSP60) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. Reactivity is specific to human samples.

The assay uses a microplate pre-coated with antibody specific for HSP60. Standards and samples are added to the wells and HSP60 is captured by the immobilized antibody. Following incubation and washing, a biotinylated anti-HSP60 antibody binds the captured antigen, after which an HRP-avidin conjugate is added. TMB substrate solution is then used to visualise the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader and HSP60 concentration is calculated by reference to the standard curve.

The measuring range is 0.156–10 ng/ml with a sensitivity of 0.063 ng/ml. Total assay time is 4 hours 30 minutes on a pre-coated microplate platform of 12 x 8 well strips, with colourimetric detection. Spike-and-recovery validation gave averages of 91% for serum (range 80–102%, n=5), 90% for EDTA plasma (range 81–100%, n=5) and 84% for heparin plasma (range 80–89%, n=5). Kit components should be stored at +4°C or -20°C as instructed and the kit must not be used past its expiration date.