**Human Galactose 6-Sulfatase (Gal-6S) ELISA Kit – Instructions for Use**
This ELISA kit is designed for the quantitative determination of Human Galactose 6-Sulfatase (Gal-6S) in various biological samples. It employs a sandwich ELISA method, which ensures high specificity and sensitivity for detecting Gal-6S levels.
**Kit Specifications:**
- **Configuration:** 48-well or 96-well format
- **Standard Dilution:** 1.5 mL × 1 vial
- **Enzyme Standard Reagent:** 3 mL × 1 vial (for 48-well) or 6 mL × 1 vial (for 96-well)
- **Storage Conditions:** All reagents should be stored at 2–8°C unless otherwise stated.
**Kit Components:**
- **Sealing Film:** 2 pieces (for 48-well) / 2 pieces (for 96-well)
- **Standard:** 0.5 mL × 1 vial, 2700 ng/mL
- **Sample Diluent:** 3 mL × 1 vial (for 48-well) or 6 mL × 1 vial (for 96-well)
- **Developer A:** 3 mL × 1 vial (for 48-well) or 6 mL × 1 vial (for 96-well)
- **Chromogen B Solution:** 3 mL × 1 vial (for 48-well) or 6 mL × 1 vial (for 96-well), store at -20°C
- **Stop Solution:** 3 mL × 1 vial (for 48-well) or 6 mL × 1 vial (for 96-well)
- **Concentrated Wash Solution:** 20 mL × 20 times (for 48-well) or 20 mL × 30 times (for 96-well), store at 2–8°C
**Principle of the Assay:**
The Human Galactose 6-Sulfatase ELISA Kit uses a double-antibody sandwich technique. The microtiter plate is pre-coated with a specific antibody against Gal-6S. After incubation with the sample, the Gal-6S present in the sample binds to the immobilized antibody. An HRP-conjugated secondary antibody is then added, forming a complex. Following a wash step, TMB substrate is introduced, which turns blue in the presence of HRP. The reaction is stopped by adding a stop solution, causing the color to shift to yellow. The intensity of the yellow color is directly proportional to the concentration of Gal-6S in the sample.
**Procedure Summary:**
1. Prepare standard and sample dilutions as specified.
2. Add standards and samples to the appropriate wells.
3. Incubate and wash the plate.
4. Add HRP-labeled detection antibody.
5. Incubate and wash again.
6. Add TMB substrate and incubate.
7. Stop the reaction with the stop solution.
8. Measure OD at 450 nm using a microplate reader.
9. Plot a standard curve using the OD values of the standards.
10. Calculate the Gal-6S concentration of the samples based on the standard curve and apply any necessary dilution factors.
**Notes:**
- This kit is intended for research use only.
- Ensure all reagents are brought to room temperature before use.
- Avoid contamination during the procedure to maintain accuracy.
- Follow all safety guidelines when handling chemicals and biological materials.
**Important:**
For best results, perform the assay within the recommended time frame and strictly follow the protocol provided. Any deviations may affect the accuracy and reproducibility of the results.
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