**Human Galactose 6-Sulfatase (Gal-6S) ELISA Kit – Instructions for Use**
**Kit Specifications:**
This ELISA kit is available in a 48-well or 96-well configuration. It is designed for the quantitative determination of Human Galactose 6-Sulfatase (Gal-6S) in biological samples such as serum, plasma, and cell culture supernatants.
**Standard Dilution:**
Each kit includes one vial of enzyme standard reagent: 1.5 ml × 1 bottle (for 48-well) or 3 ml × 1 bottle (for 96-well). The standard concentration is 2700 ng/L.
**Storage Conditions:**
All components should be stored at 2–8°C, except for the chromogen B solution, which should be stored at -20°C.
**Kit Components:**
- Sealing Film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 0.5 ml × 1 bottle
- Enzyme Standard: 1×48 / 1×96
- Sample Diluent: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Developer A: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Chromogen B: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Stop Solution: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96)
- Concentrated Wash Solution: 20 ml × 20 times (48) / 20 ml × 30 times (96)
**Principle of the Assay:**
This ELISA kit utilizes the sandwich immunoassay method. The microtiter plate is pre-coated with a specific antibody against Human Galactose 6-Sulfatase (Gal-6S). After incubation with the sample, the target antigen binds to the immobilized antibody. A HRP-conjugated secondary antibody is then added, forming a complex. Following washing steps, TMB substrate is added, and the reaction is stopped with a stop solution. The color development is directly proportional to the concentration of Gal-6S in the sample. The optical density (OD) is measured at 450 nm using a microplate reader.
**Procedure Summary:**
1. Prepare standard dilutions according to the instructions.
2. Add standards, controls, and samples to the appropriate wells.
3. Incubate and wash the plate.
4. Add HRP-labeled antibody and incubate.
5. Wash again and add TMB substrate.
6. Stop the reaction and measure OD values.
7. Plot a standard curve using the OD values of the standards.
8. Calculate the Gal-6S concentration in the samples by interpolating the OD values on the standard curve.
9. Adjust for any dilution factor if applicable.
**Note:** This kit is intended for research use only. It is not approved for diagnostic or clinical use. Always follow proper safety procedures when handling reagents and samples.
**Important Tips:**
- Ensure all reagents are brought to room temperature before use.
- Avoid contamination during the assay.
- Perform the experiment in duplicate to ensure accuracy.
- Follow the recommended incubation times and temperatures strictly.
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