**Human AGEsR ELISA Kit – For the Quantitative In Vitro Determination of Human Advanced Glycation End Products Receptor Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Biological Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
This kit is designed for the quantitative measurement of Human AGEsR (Advanced Glycation End Products Receptor) levels in various biological samples. The principle of the assay is based on a competitive enzyme-linked immunosorbent assay (ELISA), where the concentration of AGEsR in the sample is determined by comparing its optical density (OD) to a standard curve generated using a set of calibration standards.
The Stop Solution changes the color from blue to yellow, allowing for accurate quantification. The calibration standards are run alongside the samples to create a standard curve, which is used to calculate the exact concentration of AGEsR in each sample.
**Sample Collection and Storage:**
- **Serum:** Use a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifugation at 2000×g for 20 minutes. Remove the serum and assay immediately or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect plasma using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g, 2–8°C. Avoid freeze-thaw cycles.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure samples are properly centrifuged and free from hemolysis or debris.
**Materials Required but Not Supplied:**
1. Incubator set 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 strips / 12×4 strips)
- Standards (6 vials, 0.5 ml/vial)
- Sample Diluent (6.0 ml / 3.0 ml)
- HRP-Conjugate Reagent (10.0 ml / 5.0 ml)
- 20X Wash Solution (25 ml / 15 ml)
- Chromogen Solution A (6.0 ml / 3.0 ml)
- Chromogen Solution B (6.0 ml / 3.0 ml)
- Stop Solution (6.0 ml / 3.0 ml)
- Closure Plate Membrane (2 units)
- User Manual (1 copy)
- Sealed Bags (1 unit)
**Standard Concentrations:** 640, 320, 160, 80, 40, 20 pg/ml. If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
**Precautions:**
- Do not mix reagents from different kit lots.
- Allow all reagents to reach room temperature (20–25°C) before use.
- Do not use reagents past their expiration date.
- Store unused microtiter strips in sealed bags with desiccant.
- Handle all blood-derived samples with caution. Wear gloves and follow good laboratory practices.
- Dispose of waste after at least 30 minutes to inactivate viruses.
- Keep chromogen B away from heat and flame due to its acetone content.
**Reagent Preparation:**
- Wash Solution (1X): Dilute 1 volume of 20X Wash Solution with 19 volumes of distilled or deionized water. Stable for 1 month at 2–8°C.
**Assay Procedure:**
1. Prepare all reagents before starting. Run standards and samples in duplicate.
2. Add 50 μl of standard or sample to appropriate wells. Blank well contains no sample.
3. Add 100 μl of HRP-conjugate reagent to all wells except blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the plate 4 times manually or automatically.
5. Add 50 μl of Chromogen A and 50 μl of Chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
6. Add 50 μl of Stop Solution. The color should turn yellow. Gently tap if uneven.
7. Read OD at 450 nm. Calculate mean OD for each standard and sample, subtracting blank values. Construct standard curve and determine sample concentrations.
**Performance Characteristics:**
- Intra-assay and Inter-assay CV <15%
- Assay Range: 20–640 pg/mL
- Sensitivity: <1.0 pg/mL
- Cross-reactivity: No significant cross-reactivity with other proteins
- Storage: 2–8°C (frequent use); -20°C for long-term storage (up to 6 months)
**Important Notes:**
- Always read the entire package insert before use.
- Follow all safety protocols when handling biological samples.
- Each user should generate their own standard curve.
- Results may vary slightly depending on operator technique, incubation time, and temperature.
**Note:** This product is intended for research purposes only and is not approved for clinical diagnostics.
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