**Human CP IgG ELISA Kit – For the quantitative in vitro determination of Human Chlamydia psittaci IgG concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.**
Before starting the assay, please read this entire package insert carefully to ensure proper use of the kit. This ELISA kit is designed for the accurate measurement of Human Chlamydia psittaci IgG levels in various biological samples. The colorimetric reaction is measured at 450 nm using a spectrophotometer.
The kit includes a set of calibration standards that are run alongside the samples to generate a standard curve. By comparing the optical density (OD) values of the unknown samples to this curve, the concentration of CP IgG can be accurately determined.
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**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. Store at -20°C. Avoid repeated freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other body fluids:** Centrifuge to remove particulates, then assay immediately or aliquot and store at -20°C. Ensure no hemolysis or granules are present in the sample.
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**Materials Required but Not Supplied:**
1. 37°C incubator
2. Microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
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**Reagents Provided (Stored at 2–8°C):**
| Reagent Name | 96 Determinations | 48 Determinations |
|---------------------------|-------------------|-------------------|
| MicroELISA Strip Plate | 12×8 strips | 12×4 strips |
| Standard (6 vials) | 0.5 ml/vial | 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 | 2 |
| User Manual | 1 | 1 |
| Sealed Bags | 1 | 1 |
*Note: Standard concentrations are 640, 320, 160, 80, 40, 20 pg/mL.*
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**Important Instructions:**
- Do not use water baths to thaw samples or reagents.
- Never use reagents beyond their expiration date.
- Always use deionized or distilled water.
- Keep microtiter plates in sealed bags until needed. Unused strips should be stored at 2–8°C with desiccant.
- Use fresh pipette tips for each transfer to prevent cross-contamination.
- Disposable gloves must be worn during the procedure. All biological materials should be treated as potentially infectious.
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**Waste Disposal:**
- For liquid waste, add sodium hypochlorite to a final concentration of 1.0%. Let stand for at least 30 minutes before disposal.
- Substrate solution is sensitive to contamination.
- Chromogen B contains 20% acetone; keep away from heat or flame.
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**Reagent Preparation:**
- **Wash Solution (1X):** Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to 1 month.
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**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 receives no addition.
3. Add 100 µL of HRP-conjugate reagent to all wells except blank. Cover 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 changes from blue to yellow. If uneven, gently tap the plate.
7. Measure OD at 450 nm using a microplate reader.
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**Data Analysis:**
- Plot average OD values (450 nm) of standards on the Y-axis against their concentrations on the X-axis.
- Subtract blank OD from all readings.
- Locate the sample OD on the Y-axis, draw a horizontal line to the curve, and drop a vertical line to the X-axis to determine concentration.
- Intra-assay and inter-assay CVs are <15%.
- Assay range: 20 pg/mL – 640 pg/mL.
- Sensitivity: <10 pg/mL.
- Cross-reactivity: No significant interference observed.
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**Storage:**
- Store at 2–8°C for frequent use; up to 6 months at -20°C.
Always follow good laboratory practices when handling biological samples. This kit is intended for research purposes only.
Foundation Cushion
RYEOGLOBAL Co,.LTD , https://www.ryeolab.com