Human surface membrane immunoglobulin A (mIgA) Eelisa kit instruction manual
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**Human Surface Membrane Immunoglobulin A (mIgA) ELISA Kit – Instructions for Use**
**Kit Specifications:**
- 48-well or 96-well configuration
- Standard dilution: 1.5 ml × 1 vial
- Enzyme standard reagent: 3 ml × 1 vial (48-well) / 6 ml × 1 vial (96-well)
- This kit is intended for research use only.
**Preparation of Standard Curve:**
To determine the concentration of mIgA in the sample, prepare a standard curve by plotting the standard concentrations on the x-axis and the corresponding OD values on the y-axis. Alternatively, calculate the linear regression equation using the standard concentrations and their OD values. Input the sample OD value into this equation to obtain the calculated concentration, then multiply by the dilution factor to get the actual sample concentration.
**Kit Components:**
- Sealing film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 2700 ng/L, 0.5 ml × 1 vial
- Enzyme standard: 1×48 / 1×96
- 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: 3 ml × 1 vial (48-well) / 6 ml × 1 vial (96-well)
- Concentrated wash solution: 20 ml × 20 times × 1 bottle (48-well) / 20 ml × 30 times × 1 bottle (96-well)
**Storage Conditions:**
- 2–8°C for all components
- Shelf life: 6 months from the date of manufacture
**Principle of Operation:**
The kit employs a double-antibody sandwich ELISA method to quantify human surface membrane immunoglobulin A (mIgA). A purified mIgA antibody is coated onto a microplate. The sample is added, followed by an HRP-labeled mIgA antibody, forming a complex. After washing, TMB substrate is added, which turns blue under HRP activity and then yellow upon acid termination. The color intensity correlates with the mIgA concentration, measured at 450 nm.
**Purpose:**
This ELISA kit is designed to detect mIgA levels in human serum, plasma, urine, cell culture supernatants, and other biological fluids.
**Sample Preparation Guidelines:**
1. **Serum:** Allow blood to clot at room temperature, centrifuge at 2000–3000 rpm for 10–20 minutes, and collect the supernatant.
2. **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix well, centrifuge, and collect the supernatant.
3. **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes, and collect the supernatant.
4. **Cell Culture Supernatant:** Centrifuge and collect the supernatant; for intracellular components, lyse cells via freeze-thaw cycles before centrifuging.
5. **Tissue:** Homogenize in PBS, centrifuge, and collect the supernatant.
6. **Storage:** Store samples at -20°C if not tested immediately. Avoid repeated freezing and thawing.
**Important Notes:**
- Do not use samples containing NaN3, as it inhibits HRP activity.
- Ensure all reagents are brought to room temperature before use.
- Prepare a standard curve each time and run duplicates for accuracy.
- Use separate pipettes for each step to avoid cross-contamination.
- Keep the substrate away from light.
- All waste materials should be handled as biohazardous.
- Do not mix components from different batches.
- Follow the manual strictly, and rely on the microplate reader for accurate results.
**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 technical assistance is available during working hours. We also offer free sample testing services to ensure optimal experimental outcomes.
**Delivery:**
Order-to-delivery within the agreed timeframe.
**Note:** For English instructions, the English version takes precedence.