**Human FcεRII ELISA Kit – For the quantitative in vitro determination of Human Receptor II for the Fc region of Immunoglobulin E concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other biological fluids. FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.**
This ELISA kit is designed to measure the concentration of FcεRII in various biological samples. It includes a set of calibration standards that are run alongside the test samples. These standards help generate a standard curve by plotting optical density (OD) values against known FcεRII concentrations. The sample concentrations are then calculated by comparing their OD readings to this standard curve.
**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 serum and assay immediately or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g (2–8°C). Store at -20°C. Avoid repeated freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids:** Centrifuge to remove particulates. Assay immediately or aliquot and store at -20°C. Ensure no hemolysis or granules are present.
**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):**
- Microtiter Strip Plate (12×8 or 12×4 strips)
- Standard (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 Solution A (6.0 ml or 3.0 ml)
- Chromogen Solution B (6.0 ml or 3.0 ml)
- Stop Solution (6.0 ml or 3.0 ml)
- Closure Plate Membrane (2 pieces)
- User Manual (1 copy)
- Sealed Bags (1 bag)
**Notes:**
- Standard concentrations: 20, 10, 5, 2.5, 1.25, 0.625 ng/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. Use only manufacturer-supplied reagents.
- Allow all reagents and materials to reach room temperature (20–25°C) before use. Do not use water baths to thaw samples.
- Do not use beyond the expiration date.
- Only use deionized or distilled water for dilutions.
- Keep unused microtiter plate strips in the storage bag with desiccant.
- Wear disposable gloves during handling. Treat all blood-derived samples as potentially infectious.
- Dispose of samples and waste following proper viral inactivation protocols.
- Substrate solution may be contaminated; avoid using if discolored.
- Chromogen B contains 20% acetone—keep away from heat or flame.
**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 1 month.
**Assay Procedure:**
1. Prepare all reagents before starting the assay. Run each standard and sample 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 the blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the microtiter 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 change from blue to yellow. If green or uneven, gently tap the plate.
7. Measure OD at 450 nm. Plot average OD values of standards vs. concentrations to create a standard curve.
**Data Interpretation:**
- Calculate mean OD for each standard and sample.
- Subtract blank OD from all values before interpretation.
- Locate the sample OD on the Y-axis and extend a line to the standard curve to determine concentration.
- Intra-assay and inter-assay CVs are <15%.
**Assay Range:** 0.625 – 20 ng/ml
**Sensitivity:** <0.1 ng/ml
**Cross-Reactivity:** No significant cross-reactivity observed.
**Storage:** 2–8°C (frequent use); 6 months at -20°C.
**Note:** Always read the full instructions before use. This product is intended for research purposes only.
Lounge chair,High Quality Lounge chair,Lounge chair Details, CN
Queen Furniture Co., Ltd. , https://www.queenfurnitures.com