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Scientific Background
This ELISA kit applies to the in vitro Qualitative determination of SARS-CoV-2 Neutralization Antibody in serum or plasma.
Assay Principle
This ELISA kit uses Competitive-ELISA as the method to qualitatively detect the Anti-SARS-CoV-2 Neutralization Antibody in the sample. The micro ELISA plate provided in this kit is pre-coated with recombinant human ACE2. During the reaction, the SARS-CoV-2 Neutralization Antibody in the pretreated samples or controls competes with a fixed amount of human ACE2 on the solid phase supporter for sites on the Horseradish peroxidase (HRP) conjugated recombinant SARS-CoV-2 RBD fragment (HRP-RBD). After 37℃ incubation, the unbound HRP-RBD as well as any HRP-RBD bound to non-Neutralization antibody will be captured on the plate and eventually form the ACE2-RBD-HRP complex, while the circulating neutralization antibodies HRP-RBD complexes remain in the supernatant and are removed during washing. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. Compared with the inhibition ratio to judge whether SARS-CoV-2 Neutralization Antibody exists in the tested samples or not.
Performance Specifications
| Total Assay Time | 1 h 30 min |
|---|---|
| Compatible Sample Types | Serum, plasma |
| Species Reactivity | Human |
| Detection Method | Competitive |
| Precision (CV) | Both intra-CV and inter-CV are < 15%. |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Human SARS-CoV-2 Neutralization Antibody in samples.No significant cross-reactivity or interference between Human SARS-CoV-2 Neutralization Antibody and analogues was observed
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with Serum, plasma. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is specified in the kit datasheet. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 1 h 30 min, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Competitive ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant SARS-CoV-2 Neutralization Antibody standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
Tissue samples may contain endogenous or exogenous proteases during processing, leading to degradation of extracted proteins. Therefore, it's necessary to add protease inhibitors during processing to ensure the integrity of target proteins. If customers can keep samples cold and handle them quickly during processing, omitting the protease inhibitor may not have a significant effect. After preparation, samples should be tested promptly or immediately aliquoted and frozen at -20°C or -80°C.
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