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Scientific Background
This ELISA kit applies to the in vitro Qualitative determination of SARS-CoV-2 IgM in serum or plasma.
Assay Principle
This ELISA kit uses Indirect-ELISA as the method to qualitatively detect the SARS-CoV-2 Nucleocapsid protein IgM in the sample. The micro ELISA plate provided in this kit is pre-coated with purified SARS-CoV-2 Nucleocapsid protein (E-ELC-001) antigen, after adding samples to wells, the SARS-CoV-2 Nucleocapsid protein IgM in the samples will combine with the pre-coated SARS-CoV-2 Nucleocapsid protein antigen. After washing completely, add Horseradish Peroxidase (HRP) conjugated mouse anti human IgM to develop the antigen-antibody-HRP conjugated secondary antibody complex. Free components are washed away, then the substrate solution is added to each well. Only those wells that contain SARS-CoV-2 Nucleocapsid protein IgM and HRP conjugated anti-human IgM will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. Compared with the CUT OFF value to judge whether SARS-CoV-2 Nucleocapsid protein IgM 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 | Sandwich |
| Precision (CV) | Intra-CV: CV%＜8%; Inter-CV: CV%＜10% |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Human SARS-CoV-2 IgM in samples.No significant cross-reactivity or interference between Human SARS-CoV-2 IgM 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 Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant SARS-CoV-2 IgM 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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