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Kaixin micro test
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Test probe P100-M3
**Human Interferon Beta (IFN-β/IFNB) ELISA Kit Instruction Manual**
**ElisaKit Specifications:**
Available in 48-well or 96-well configurations.
**Standard Dilution:**
1.5ml × 1 bottle
**Enzyme Standard Reagent:**
3ml × 1 bottle (for 48-well) / 6ml × 1 bottle (for 96-well)
**Note:** This reagent is for research use only.
**Calculation Method:**
Plot the standard concentrations on the x-axis and OD values on the y-axis to create a standard curve. Determine the sample concentration from the curve based on its OD value, then multiply by the dilution factor. Alternatively, calculate the linear regression equation using the standard data and substitute the sample's OD into the equation for accurate results.
**Kit Composition:**
- Sealing film: 2 pieces (48) / 2 pieces (96)
- Instructions: 1 copy
- Sealed bag: 1
- Standard: 2700ng/L, 0.5ml × 1 bottle, stored at 2–8°C
- Enzyme-labeled plate: 1×48 or 1×96, stored at 2–8°C
- Sample diluent: 3ml × 1 bottle (48) / 6ml × 1 bottle (96), stored at 2–8°C
- Reagent A: 3ml × 1 bottle (48) / 6ml × 1 bottle (96), stored at 2–8°C
- Developer B solution: 3ml × 1 bottle (48) / 6ml × 1 bottle (96), stored at 2–8°C
- Stop solution: 3ml × 1 bottle (48) / 6ml × 1 bottle (96), stored at 2–8°C
- Concentrated washing solution: (20ml × 20 times) × 1 bottle (for 48) or (20ml × 30 times) × 1 bottle (for 96), stored at 2–8°C
**Experimental Principle:**
This kit uses a double-antibody sandwich ELISA method to detect human IFN-β levels. The microwell plate is coated with purified IFN-β antibody. After adding the sample, the HRP-labeled IFN-β antibody binds to form a complex. After washing, TMB substrate is added, which turns blue under HRP catalysis and then yellow when acid is added. The color intensity is directly proportional to the IFN-β concentration. The absorbance is measured at 450nm, and the concentration is calculated from the standard curve.
**Purpose:**
To determine the concentration of interferon-beta (IFN-β/IFNB) in human serum, plasma, urine, cell culture supernatants, tissue homogenates, and other biological fluids.
**Guarantee & Support:**
We provide free technical support during working hours. Please contact us for assistance.
We also offer free sample testing services to help you achieve optimal experimental results.
**Storage Conditions & Expiry:**
- Storage: 2–8°C
- Shelf Life: 6 months
**Sample Preparation Guidelines:**
1. **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes.
2. **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix for 10–20 minutes, then centrifuge.
3. **Urine:** Collect with a sterile tube, centrifuge, and collect supernatant.
4. **Cell Culture Supernatant:** Centrifuge after collection; for intracellular components, lyse cells via freezing/thawing.
5. **Tissue:** Homogenize in PBS, centrifuge, and collect supernatant.
6. **General:** Process samples as soon as possible after collection. If not tested immediately, store at –20°C, avoiding repeated freeze-thaw cycles.
7. **Avoid:** Samples containing NaN3, as it inhibits HRP activity.
**Step-by-Step Procedure:**
1. **Standard Dilution:** Prepare 10 standards in serial dilutions (1800 ng/L to 150 ng/L).
2. **Loading:** Add 40 μl sample diluent and 10 μl sample to each well (final 5x dilution).
3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes.
4. **Washing:** Dilute concentrated wash solution and wash 5 times.
5. **Add Enzyme:** Add 50 μl enzyme reagent to each well except blank.
6. **Incubation:** Repeat incubation at 37°C for 30 minutes.
7. **Color Development:** Add 50 μl developer and incubate at 37°C for 15 minutes.
8. **Stop Reaction:** Add 50 μl stop solution to each well.
9. **Measurement:** Read OD at 450 nm within 15 minutes of stopping the reaction.
**Important Notes:**
1. Let the kit equilibrate to room temperature before use. Store unopened plates in a sealed bag.
2. Crystallization in washing solution is normal; dissolve if necessary.
3. Use accurate pipettes and avoid cross-contamination.
4. Always run a standard curve and consider diluting high-concentration samples.
5. Use a new sealing film for each experiment.
6. Keep substrates away from light.
7. Follow instructions strictly and rely on microplate reader readings.
8. Treat all waste as biohazardous materials.
9. Do not mix reagents from different batches.
10. In case of discrepancies, the English manual takes precedence.
**Performance:**
- Correlation coefficient (R) ≥ 0.95
- Intra-batch variation < 9%, Inter-batch variation < 11%
**Detection Range:**
0.2 IU/L – 6 IU/L
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