**Human Chlamydia psittaci 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.**
Please read this package insert carefully before use. The ELISA method is a widely used technique for detecting and quantifying specific antibodies or antigens in biological samples. In this kit, the color development is measured at 450 nm using a spectrophotometer. To determine the concentration of CP IgG in the sample, the kit includes a set of calibration standards. These standards are run alongside the samples to generate a standard curve, which is essential for accurately determining the IgG levels in unknown samples.
The optical density (OD) values obtained from the samples are compared to the standard curve to calculate the corresponding CP IgG concentrations. This process ensures reliable and reproducible results when performed correctly.
**Sample Collection and Storage:**
- **Serum:** Use a serum separator tube and allow the blood to clot for 2 hours at room temperature or overnight at 4°C. Centrifuge at approximately 2000×g for 20 minutes. Store samples at -20°C. Avoid repeated freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids:** Remove particulates by centrifugation, and assay immediately or aliquot and store at -20°C. Ensure proper centrifugation and avoid hemolysis or granulation.
**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
**Reagents Provided:**
All reagents should be stored at 2–8°C. Check the expiration date on the label before use.
| Name | 96 Determinations | 48 Determinations |
|------------------------------|-------------------|-------------------|
| MicroELISA Stripplate | 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 |
**Important Notes:**
1. The standard concentrations are 640, 320, 160, 80, 40, and 20 pg/mL.
2. If sample values exceed the highest standard, dilute with Sample Diluent.
3. Do not use water baths to thaw samples or reagents.
4. Do not use kit components past their expiration date.
5. Only use deionized or distilled water.
6. Do not remove microtiter plates from storage bags until needed. Unused strips should be kept at 2–8°C in sealed pouches with desiccant.
7. Use fresh pipette tips for each transfer to prevent cross-contamination.
8. Disposable gloves must be worn during the procedure. All blood-derived products should be considered potentially infectious.
9. Dispose of all samples and waste according to local regulations.
10. Liquid waste should be treated with sodium hypochlorite (final concentration 1.0%) for at least 30 minutes before disposal.
11. Substrate solutions are sensitive to contamination. Avoid exposure to light.
12. Chromogen B contains 20% acetone—keep away from heat or flame.
13. Allow all reagents to reach room temperature (20–25°C) before use.
**Reagent Preparation and Storage:**
- **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 one month.
**Assay Procedure:**
1. Prepare all reagents before starting.
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 the blank. Cover with an adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the microtiter plate four times manually or automatically. Ensure complete removal of liquid after each wash.
5. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protecting from light.
6. Add 50 µL of Stop Solution to each well. The color should turn yellow; if green, gently mix.
7. Read OD at 450 nm using a microplate reader.
**Data Interpretation:**
- Generate a standard curve by plotting average OD values (450 nm) against standard concentrations.
- Subtract blank OD from all readings before interpretation.
- Locate the sample OD on the Y-axis and draw a horizontal line to intersect the standard curve. Then draw a vertical line to the X-axis to find the corresponding concentration.
- Each user should generate their own standard curve due to potential variability in technique, timing, or environmental conditions.
- Intra-assay and inter-assay CV% are less than 15%.
- Assay range: 20 pg/mL to 640 pg/mL.
- Sensitivity: <10 pg/mL.
- Cross-reactivity: No significant cross-reactivity observed.
- Storage: 2–8°C (frequent use); 6 months at -20°C.
**Safety and Handling:**
- Always follow good laboratory practices.
- Treat all biological materials as potentially infectious.
- Wear appropriate personal protective equipment (PPE) throughout the procedure.
**Note:** This product is intended for research purposes only and is not approved for clinical diagnostic use.
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