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 reagent is intended solely for research purposes. To determine the concentration of mIgA E in your samples, plot a standard curve using the standard concentrations on the x-axis and OD values on the y-axis. Alternatively, calculate the linear regression equation based on the standard curve and use it to determine the sample concentration from its OD value. Multiply by the dilution factor to obtain the actual concentration.
**Kit Composition:**
- Sealing film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 2700 ng/L, 0.5 ml × 1 vial
- Enzyme standard package: 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 solution: 3 ml × 1 vial (48-well) / 6 ml × 1 vial (96-well)
- Concentrated wash solution: 20 ml × 20 times (48-well) / 20 ml × 30 times (96-well)
**Storage Conditions:**
- Store all components at 2–8°C
- Shelf life: 6 months from the date of manufacture
**Principle of the Assay:**
The kit uses a double-antibody sandwich ELISA method to detect human surface membrane immunoglobulin A (mIgA) E in the sample. The microplate is pre-coated with a purified anti-mIgA E antibody. After adding the sample, the target antigen binds to the immobilized antibody. Then, an HRP-conjugated secondary antibody is added, forming a complex. After washing, TMB substrate is added, and the reaction is stopped. The color intensity is proportional to the amount of mIgA E in the sample. The absorbance is measured at 450 nm, and the concentration is determined using a standard curve.
**Purpose:**
This ELISA kit is designed for the quantitative determination of mIgA E in human serum, plasma, urine, cell culture supernatants, and other biological fluids.
**Sample Preparation Guidelines:**
1. **Serum:** Allow blood to clot at room temperature, then centrifuge at 2000–3000 rpm for 10–20 minutes.
2. **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix well, and centrifuge similarly.
3. **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
4. **Cell culture supernatant:** Centrifuge after collection; for intracellular components, lyse cells by freezing/thawing.
5. **Tissue:** Homogenize in PBS, centrifuge, and collect the supernatant.
6. **Storage:** Samples should be processed as soon as possible. If not tested immediately, store at -20°C, avoiding repeated freeze-thaw cycles.
**Notes:**
- Avoid using samples containing NaN3, as it may inhibit HRP activity.
- Ensure proper equilibration of the kit before use.
- Always prepare a standard curve with duplicates for accurate results.
- Handle all reagents carefully, and avoid cross-contamination.
- Keep the substrate away from light during the reaction.
- Follow the manual strictly, and rely on microplate reader readings for final results.
- All waste materials should be treated as biohazardous.
**Performance Characteristics:**
- Correlation coefficient (R²) ≥ 0.95
- Intra-assay and inter-assay variation < 9% and < 11%, respectively
- Detection range: 0.2 IU/L – 6 IU/L
**Service Commitment:**
- Free technical support during working hours
- Free sample testing services available upon request
- Fast delivery after payment confirmation
This kit is a reliable tool for researchers studying immune responses and related biomarkers. Proper handling and adherence to instructions are essential for obtaining accurate and reproducible results.