This human Cathepsin D BioAssay™ ELISA Kit is based on the standard sandwich enzyme-linked immunosorbent assay technology.
Sample Type
Serum and plasma (heparin, EDTA), cell culture supernatants.
Intended Use
For use in quantitative protein detection, establishing normal range etc.
Specificity
Recognizes natural and recombinant human Cathepsin D. This ELISA kit shows no cross-reactivity with other relevant proteins.
Test Principle
This assay employs a monoclonal antibody form mouse specific for Cathepsin D coated on a 96-well plate. Standards (NSO) and test samples are added to the wells and Cathepsin D present in a sample is bound to the wells by the immobilized antibody. A biotinylated detection polyclonal antibody from goat specific for Cathepsin D is added subsequently. After washing away the unbound biotinylated antibody with PBS or TBS buffer, Avidin-Biotin-Peroxidase Complex is added to the wells. The wells are again washed with PBS or TBS buffer to remove the unbound conjugates. HRP substrate TMB is used to visualize the HRP enzymatic reaction. TMB is catalyzed by HRP to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow color is proportional to the human Cathepsin D captured onto the plate.
Kit Components
Microplate coated with anti-human Ab against Cathepsin D, 1x96 wellsLyophilized recombinant human Cathepsin D standard (10ng/vial), 2x10ng Biotinylated anti-human Cathepsin D antibody, 1x130ulAvidin-Biotin-Peroxidase Complex (ABC), 1x130ulSample diluent buffer, 1x30mlAntibody diluent buffer, 1x12mlABC diluent buffer, 1x12mlTMB color developing agent (Colorless), 1x10mlTMB stop solution, 1x10ml
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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