The Human Heat Shock Protein 70 (HSP70) ELISA Kit allows for the in vitro quantitative determination of HSP70 concentrations in serum, plasma, or tissue homogenates. This assay kit was designed and optimized for neuroscience research use in humans. It has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details.
This assay employs the quantitative sandwich enzyme immunoassay technique, in which HSP70 in the samples or standards are sandwiched between pre-coated HSP70 antibody and Biotin-conjugated HSP70 antibody. HRP-avidin is then added into the wells. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of HSP70 bound in the initial step. The color development is stopped upon adding the stop solution, and the intensity of the color is measured at 450 nm via a microplate reader. The levels of HSP70 in the samples can be determined by referring to the O.D. (optical density) of the samples to the standard curve.
HSP70 is generated in response to stress or constitutively as the major chaperone contributing to maintaining protein homeostasis in both eukaryotes and prokaryotes. HSP70 binds to client proteins in the early stages of protein folding, which controls the availability of such regions, promoting the formation of the proper protein fold, but inhibiting aggregate formation. Upon detection of incorrect protein folding, HSP70 associates with additional co-chaperones to facilitate degradation of the misfolded protein. It also enhances bacterial survival in hostile environments.