**Human Anti-Nuclear Antibody (ANA) ELISA Kit Instruction Manual**
**Kit Specifications:**
This Human Anti-Nuclear Antibody (ANA) ELISA Kit is available in 48-well or 96-well configurations. The kit includes all necessary reagents for quantitative detection of ANA in human serum or plasma samples.
**Standard Dilution:**
1.5 mL × 1 vial of enzyme standard reagent.
- For 48-well configuration: 3 mL × 1 vial
- For 96-well configuration: 6 mL × 1 vial
**Calculation Method:**
To determine the concentration of ANA in your sample, plot a standard curve using the concentration of the standard on the x-axis and the corresponding OD values on the y-axis. Draw the standard curve on graph paper, then use the OD value of your sample to estimate its concentration from the curve. Multiply this value by the dilution factor to obtain the actual concentration of the sample. Alternatively, calculate the linear regression equation from the standard curve data and substitute the sample’s OD value into the equation to directly compute the concentration. Don’t forget to multiply by the dilution factor for accurate results.
**Kit Composition:**
- Sealing Film: 2 pieces (for 48-well), 2 pieces (for 96-well)
- Standard: 2700 ng/L, 0.5 mL × 1 vial
- Enzyme Standard: 1×48, 1×96 (stored at 2–8°C)
- Sample Diluent: 3 mL × 1 vial (for 48-well), 6 mL × 1 vial (for 96-well)
- Developer A: 3 mL × 1 vial (for 48-well), 6 mL × 1 vial (for 96-well)
- Chromogen B: 3 mL × 1 vial (for 48-well), 6 mL × 1 vial (for 96-well) – store at -20°C
- Stop Solution: 3 mL × 1 vial (for 48-well), 6 mL × 1 vial (for 96-well)
- Concentrated Wash Solution: 20 mL × 20 times (for 48-well), 20 mL × 30 times (for 96-well) – store at 2–8°C
**Storage Conditions:**
All reagents should be stored at 2–8°C unless otherwise specified. Developer A and Chromogen B must be kept at -20°C for long-term stability.
**Precautions:**
This kit is intended for research use only. Avoid contact with skin and eyes. Use appropriate personal protective equipment (PPE) when handling reagents. Ensure all measurements are performed under controlled conditions to maintain accuracy.
**Notes:**
Always check the expiration date before use. Do not mix reagents from different batches. Follow the protocol carefully to ensure reliable and reproducible results. If any step is unclear, refer to the detailed instructions provided in the manual or contact technical support for assistance.
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