**Human Anti-Epidemic Hemorrhagic Fever Virus Antibody IgM (EHF) ELISA Kit Instruction Manual**
**Kit Specifications:**
- **Configuration:** 48-well or 96-well plate
- **Standard Dilution:** 1.5 mL × 1 vial
- **Enzyme Standard Reagent:** 3 mL × 1 vial (for 48-well) / 6 mL × 1 vial (for 96-well)
- **Storage Conditions:** 2–8°C
- **Expiration:** 6 months from the date of receipt
**Purpose:**
This ELISA kit is designed for the quantitative determination of human anti-Epidemic Hemorrhagic Fever Virus Antibody IgM (EHF) in serum, plasma, urine, and other biological fluids. It is intended for research use only.
**Principle of Operation:**
The kit utilizes a sandwich ELISA method. A solid-phase antibody specific to EHF IgM is coated on microtiter wells. The sample is added, allowing the presence of EHF IgM to bind to the coated antibody. An HRP-conjugated secondary antibody is then introduced, forming a complex. After washing, TMB substrate is added, producing a color change that correlates with the concentration of EHF IgM in the sample. The reaction is stopped, and the absorbance is measured at 450 nm using a microplate reader. A standard curve is generated using known concentrations, and the sample concentration is calculated accordingly.
**Kit Components:**
- Sealing Film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 0.5 mL × 1 vial, 2700 ng/L
- Enzyme Standard: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well)
- Sample Diluent: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well)
- Developer A: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well)
- Chromogen B: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well)
- Wash Buffer: 20 mL × 20 times (48-well) / 20 mL × 30 times (96-well)
- Storage: All components should be stored at 2–8°C
**Sample Preparation Guidelines:**
- **Serum:** Allow blood to clot at room temperature, centrifuge at 2000–3000 rpm for 10 minutes. Store at -20°C if not used immediately.
- **Plasma:** Use EDTA or sodium citrate as anticoagulant. Centrifuge after mixing for 10–20 minutes.
- **Urine:** Collect in a sterile container and centrifuge for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Tissue Homogenate:** Weigh tissue, add PBS, homogenize, centrifuge, and collect supernatant.
- **Note:** Avoid repeated freeze-thaw cycles. Do not use samples containing NaN3 due to enzyme inhibition.
**Operating Procedure:**
1. **Standard Dilution:** Prepare serial dilutions of the standard solution in the microplate.
2. **Sample Loading:** Add 40 µL of sample diluent followed by 10 µL of sample to each well.
3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes.
4. **Washing:** Rinse wells 5 times with diluted wash buffer.
5. **Enzyme Addition:** Add 50 µL of enzyme-labeled reagent to each well (except blank).
6. **Second Incubation:** Repeat incubation step.
7. **Color Development:** Add 50 µL of TMB A and B, incubate at 37°C for 15 minutes.
8. **Stop Reaction:** Add 50 µL of stop solution to each well.
9. **Reading:** Measure OD at 450 nm within 15 minutes of stopping the reaction.
**Notes & Precautions:**
- Allow the kit to reach room temperature before use.
- Avoid cross-contamination by using a new sealing film for each test.
- Keep all reagents away from light.
- Always prepare a standard curve and run duplicates for accuracy.
- Follow instructions strictly; results must be based on instrument readings.
- Dispose of all waste as biohazardous material.
- Do not mix reagents from different batches.
- In case of conflict, the English manual takes precedence.
**Performance Characteristics:**
- **Sensitivity:** 0.2 IU/L
- **Dynamic Range:** 0.2–6 IU/L
- **Correlation Coefficient (R²):** ≥ 0.95
- **Intra-batch CV:** < 9%
- **Inter-batch CV:** < 11%
**Service Commitment:**
We offer free technical support during working hours. If needed, we can assist with sample testing to ensure optimal results. Our delivery time starts from payment confirmation.
**Storage Conditions:**
Store the unopened kit at 2–8°C. Once opened, store unused components in a sealed bag at 2–8°C.
**Important:** This product is for research use only and not for diagnostic purposes.
printing
Printing,Printing Anti Slip Pads,Colored Anti-Skid Mat,Anti Slip Drawer Liners
Rongxin New Material (Jiangsu) Co., Ltd. , https://www.rongxinxcljs.com