Human AHSG ELISA Kit
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**Human AHSG ELISA Kit – For the Quantitative In Vitro Determination of Human Alpha-2-HS-Glycoprotein Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
This ELISA kit is designed for the accurate quantification of Human Alpha-2-HS-Glycoprotein (AHSG) in various biological samples. The principle of the assay is based on a sandwich ELISA format, where the captured antigen binds to specific antibodies coated on the microtiter plate. A horseradish peroxidase (HRP)-conjugated secondary antibody is then added, followed by the addition of a chromogenic substrate. The reaction is stopped, and the color intensity is measured at 450 nm using a spectrophotometer. The optical density (OD) values are compared against a standard curve to determine the concentration of AHSG in the test samples.
**Intended Use:**
The Human AHSG ELISA Kit is intended for laboratory research purposes only. It is not approved for use in diagnostic or therapeutic procedures. The results obtained should not be used as a substitute for clinical diagnosis or treatment decisions.
**Sample Collection and Storage:**
- **Serum:** Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C before centrifugation at 2000×g for 20 minutes. Store aliquots at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g for 30 minutes at 2–8°C. Store at -20°C.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Centrifuge to remove particulates and store at -20°C. Ensure no hemolysis or contamination occurs.
**Materials Required but Not Supplied:**
- 37°C incubator
- Microplate reader capable of measuring absorbance at 450 nm
- Precision pipettes, disposable tips, and absorbent paper
- Distilled or deionized water
**Reagents Provided (Stored at 2–8°C):**
- 96-well microplate (12×8 strips)
- 6 standard vials (0.5 ml/vial)
- Sample diluent (6.0 ml)
- HRP-conjugate reagent (10.0 ml)
- 20X Wash solution (25 ml)
- Chromogen solutions A & B (6.0 ml each)
- Stop solution (6.0 ml)
- Closure plate membrane (2 pieces)
- User manual and sealed bags
**Standard Curve Preparation:**
Standards are prepared at concentrations of 800, 400, 200, 100, 50, and 25 μg/mL. If sample values exceed the highest standard, dilute with sample diluent and repeat the assay.
**Important Notes:**
- Do not use reagents beyond their expiration date.
- Allow all reagents to reach room temperature (20–25°C) before use.
- Avoid using water baths for thawing.
- Always use fresh pipette tips to prevent cross-contamination.
- Handle all biological materials with care, following proper biosafety protocols.
**Assay Procedure Summary:**
1. Prepare all reagents and add 50 µL of standards or samples to appropriate wells.
2. Incubate for 60 minutes at 37°C.
3. Wash the plate 4 times using 1X wash solution.
4. Add 50 µL of chromogen A and B, incubate for 15 minutes.
5. Add 50 µL of stop solution and measure OD at 450 nm.
6. Plot the standard curve and calculate sample concentrations.
**Performance Characteristics:**
- Intra-assay and inter-assay CV% < 15%
- Assay range: 25–800 μg/mL
- Sensitivity: <10 μg/mL
- Cross-reactivity: No significant cross-reaction with other proteins
**Storage Conditions:**
- 2–8°C for frequent use; up to 6 months at -20°C
**Safety and Disposal:**
All samples and reagents must be handled according to local safety regulations. Dispose of waste in accordance with biohazard guidelines. Allow liquid to stand for 30 minutes to inactivate viruses before disposal.
**Note:** This kit is not intended for use in humans or animals for medical purposes. Always follow the manufacturer’s instructions and perform tests under controlled laboratory conditions.