**Chicken Infectious Rhinitis Antibody (IC) ELISA Kit – User Manual**
**Kit Specifications:**
- 48-well or 96-well configuration
- Standard Diluent: 1.5 mL × 1 bottle
- Enzyme Standard Reagent: 3 mL × 1 bottle (for 48-well) / 6 mL × 1 bottle (for 96-well)
- This reagent is for **research use only**.
**Preparation of Standard Curve:**
To determine the concentration of the sample, plot a standard curve by using the standard concentrations on the x-axis and OD values on the y-axis. Alternatively, calculate the linear regression equation from the standard data and use it to determine the sample concentration based on its OD value. Multiply the result by the dilution factor to obtain the actual concentration.
**Kit Components:**
- Sealing Film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 2700 ng/L, 0.5 mL × 1 bottle
- Sample Diluent: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Developer A: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Chromogen B: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Stop Solution: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Concentrated Washing Solution: 20 mL × 20 times (48-well) / 20 mL × 30 times (96-well)
**Storage Conditions & Shelf Life:**
- Storage: 2–8°C
- Shelf Life: 6 months from date of manufacture
**Principle of the Assay:**
This kit uses the **double-antibody sandwich method** to detect chicken infectious rhinitis antibody (IC). The microplate is coated with purified chicken infectious rhinitis antibody (IC), and the sample is added. After incubation, HRP-labeled antibody binds to the antigen, forming an immune complex. TMB substrate is then added, resulting in a color change that correlates with the amount of antibody present. The reaction is stopped, and absorbance is measured at 450 nm.
**Purpose:**
The kit is used to quantify infectious rhinitis antibodies (IC) in chicken serum, plasma, urine, cell culture supernatants, and other biological fluids.
**Sample Preparation Guidelines:**
- **Serum:** Allow blood to clot at room temperature for 10–20 minutes, centrifuge at 2000–3000 rpm for 20 minutes.
- **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix, and centrifuge.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge after collection. For intracellular components, lyse cells via freezing/thawing and centrifuge.
- **Tissue Specimens:** Homogenize in PBS, centrifuge, and collect supernatant. Store at 2–8°C or -20°C if not tested immediately. Avoid repeated freeze-thaw cycles.
**Important Notes:**
- Do not use samples containing NaN3, as it inhibits HRP activity.
- Ensure all reagents are at room temperature before use.
- Prepare a standard curve for each test; use duplicate wells if possible.
- Avoid cross-contamination by using a new sealing film for each experiment.
- Keep substrates away from light.
- Follow the manual strictly. Results must be confirmed with a microplate reader.
- Treat all waste materials as biohazardous.
**Performance Characteristics:**
- Correlation coefficient (R²) ≥ 0.95
- Intra-assay CV < 9%, Inter-assay CV < 11%
- Detection range: 0.2 IU/L – 6 IU/L
**Technical Support:**
Free guidance and support available during working hours. We also offer free sample testing to help you achieve optimal results.
**Delivery:**
Prompt delivery upon payment. Contact us for assistance with your ELISA experiments.
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