This reagent is intended for research use only. The kit is designed to quantitatively determine the level of human Klotho protein in human serum, plasma, and other related biological samples. Principle of the assay: The kit employs a double-antibody sandwich ELISA method to detect Klotho protein. A microtiter plate is pre-coated with purified monoclonal antibodies specific to human Klotho. After adding the sample, the Klotho protein binds to the immobilized antibody on the plate. Then, a horseradish peroxidase (HRP)-labeled secondary antibody is added, forming a complex of antibody-antigen-antibody. Following thorough washing to remove unbound components, a TMB substrate solution is introduced. Under the catalytic action of HRP, TMB turns blue and then changes to yellow upon acid termination. The intensity of the color is directly proportional to the concentration of Klotho protein in the sample. The optical density (OD) is measured at 450 nm using a microplate reader, and the Klotho protein concentration is determined by comparing the OD values to a standard curve.
Kit Components:
- 48-well configuration: 1 × 48 enzyme-labeled plate, 1 × 1 bottle of standard (0.5 ml), 1 × 1 bottle of standard diluent (1.5 ml), 1 × 1 bottle of sample diluent (3 ml), 1 × 1 bottle of TMB A (3 ml), 1 × 1 bottle of TMB B (3 ml), 1 × 1 bottle of stop solution (3 ml), 1 × 1 bottle of 20× wash solution (20 ml), 2 sealing films.
- 96-well configuration: 1 × 96 enzyme-labeled plate, 1 × 1 bottle of standard (0.5 ml), 1 × 1 bottle of standard diluent (1.5 ml), 1 × 1 bottle of sample diluent (3 ml), 1 × 1 bottle of TMB A (3 ml), 1 × 1 bottle of TMB B (3 ml), 1 × 1 bottle of stop solution (3 ml), 1 × 1 bottle of 20× wash solution (23 ml), 2 sealing films.
Storage Instructions: All components should be stored at 2–8°C. The standard is stable for 180U/L. Ensure proper handling and avoid contamination during the experiment. This kit is ideal for researchers conducting studies on Klotho protein expression and its potential role in various physiological and pathological conditions.
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