**Vaspin Kit – Visceral Fat-Specific Serine Protease Inhibitor Assay**
**Purpose of the Experiment:**
This kit is designed for the quantitative determination of visceral fat-specific serine protease inhibitor (VASPIN) in human serum, plasma, and other biological fluid samples. It provides a reliable and sensitive method for measuring VASPIN levels, which are associated with metabolic health and obesity-related conditions.
**Experimental Principle:**
The VASPIN assay is based on the double-antibody sandwich ELISA technique. A microplate is pre-coated with a specific monoclonal antibody against human VASPIN. After incubation with the sample, VASPIN binds to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody is then added, forming a complex of antibody-antigen-enzyme-labeled antibody. Following a washing step, the substrate TMB (3,3',5,5'-tetramethylbenzidine) is introduced. Under HRP catalysis, TMB turns blue and then changes to yellow upon acid termination. The intensity of the color is directly proportional to the concentration of VASPIN in the sample. The optical density (OD) is measured at 450 nm using a microplate reader, and the VASPIN concentration is calculated by comparing the OD values to a standard curve.
**Kit Components:**
The kit is available in 48-well and 96-well configurations. Each configuration includes the following components:
- **Coated Plate:** 1 × 48 or 1 × 96 wells, pre-coated with anti-VASPIN antibody
- **Standard:** 0.5 ml × 1 bottle (36 ng/ml), for constructing a standard curve
- **Standard Diluent:** 1.5 ml × 1 bottle, for diluting the standard solution
- **Enzyme Conjugate:** 3 ml × 1 bottle, HRP-labeled anti-VASPIN antibody
- **Sample Diluent:** 3 ml × 1 bottle, for diluting the test samples
- **TMB Substrate Solution (A & B):** 3 ml × 1 bottle each, for color development
- **Stop Solution:** 3 ml × 1 bottle, to terminate the reaction
- **Washing Buffer (Concentrated):** 20 ml × 20 times or 20 ml × 30 times, depending on the configuration, for plate washing
**Storage Instructions:**
All reagents should be stored at 2–8°C. The coated plate should be kept sealed in its original packaging until use. Avoid repeated freeze-thaw cycles to maintain stability and performance.
**Notes:**
Proper handling and storage are essential to ensure accurate results. Always follow the manufacturer’s instructions carefully and use fresh reagents for best performance. This kit is intended for research use only and not for diagnostic purposes.
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