Human AHSG ELISA Kit
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**Human AHSG ELISA Kit – For the Quantitative In Vitro Determination of Human Alpha-2-HS-Glycoprotein in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not Intended for Diagnostic or Therapeutic Purposes.*
This ELISA kit is designed for the quantitative measurement of Human Alpha-2-HS-Glycoprotein (AHSG) in various biological samples. The assay is based on the principle of a sandwich immunoassay, where the target antigen binds to specific antibodies coated on the microtiter plate. A horseradish peroxidase (HRP)-conjugated secondary antibody is then added, followed by a chromogenic substrate that produces a color change. The reaction is stopped with a stop solution, and the optical density (OD) is measured at 450 nm using a spectrophotometer.
To determine the concentration of AHSG in the sample, a set of calibration standards is included in the kit. These standards are run alongside the samples, allowing the operator to generate a standard curve by plotting OD values against known concentrations. The sample concentration is then calculated by comparing its OD value to the standard curve.
**Sample Collection and Storage:**
- **Serum:** Use serum separator tubes, allow clotting for 2 hours at room temperature or overnight at 4°C, then centrifuge at 2000×g for 20 minutes. Store at -20°C, avoiding repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g, store at -20°C.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Remove particulates by centrifugation, assay immediately, or store at -20°C. Ensure no hemolysis or debris is present.
**Materials Required (Not Included):**
1. Incubator at 37°C
2. Microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
**Reagents Provided (Stored at 2–8°C):**
- Microtiter Strip Plate (12×8 or 12×4 strips)
- Standards (6 vials, 0.5 ml/vial)
- Sample Diluent (6.0 ml or 3.0 ml)
- HRP-Conjugate Reagent (10.0 ml or 5.0 ml)
- 20X Wash Solution (25 ml or 15 ml)
- Chromogen Solutions A & B (6.0 ml each)
- Stop Solution (6.0 ml)
- Closure Plate Membrane (2 pieces)
- User Manual and Sealed Bags
**Important Notes:**
- Standard concentrations: 800, 400, 200, 100, 50, 25 μg/mL.
- If sample values exceed the highest standard, dilute with Sample Diluent and re-run.
- Do not use reagents beyond their expiration date.
- All reagents must be brought to room temperature (20–25°C) before use.
- Avoid contamination by using fresh pipette tips for each transfer.
- Handle all samples with care, considering them potentially infectious.
**Assay Procedure Summary:**
1. Prepare all reagents and add 50 µl of standard or sample to appropriate wells.
2. Incubate for 60 minutes at 37°C, then wash 4 times manually or automatically.
3. Add 50 µl of Chromogen A and B, incubate for 15 minutes at 37°C, protected from light.
4. Add 50 µl Stop Solution and measure OD at 450 nm.
5. Plot standard curve and calculate sample concentrations.
**Performance Characteristics:**
- Sensitivity: <10 µg/mL
- Assay Range: 25–800 µg/mL
- Intra-assay CV: <15%
- Inter-assay CV: <15%
- Cross-reactivity: No significant cross-reactivity observed
**Storage Conditions:**
- 2–8°C for frequent use; up to 6 months at -20°C.
**Safety and Disposal:**
- Follow proper biohazard protocols when handling samples.
- Allow liquid to stand for 30 minutes before disposal to inactivate viruses.
- Keep Substrate B away from heat or flame due to acetone content.
This kit provides a reliable and reproducible method for quantifying AHSG levels in research settings. Always follow the manufacturer’s instructions carefully to ensure accurate results.