| Description |
CUSABIO's Human SARS-CoV-2 nucleoprotein (N) IgG Antibody ELISA Kit allows for in vitro qualitative detection of the IgG antibody-specific for SARS-CoV-2 N protein in human serum and plasma. This research-use only kit can be applied in the studies associated with COVID-19, infectious and severe pneumonia caused by SARS-CoV-2. It has been validated with high sensitivity, high specificity, and high precision.
Nucleoprotein is the most abundant protein in SARS-CoV-2. It packs the positive-sense single-stranded viral RNA into a helical ribonucleocapsid (RNP), forming the viral nucleocapsid (N). The N protein plays an important role in improving the efficiency of virus transcription and assembly. It is also involved in viral pathogenesis. N protein possessing an immunogenic effect makes it trigger an immune response, which leads to the production of IgM/G against SARA-CoV-2 N protein. SARS-CoV-2 N IgG antibodies appear in the serum after 2-3 weeks after the onset of the symptoms, peaks within the third week, and gradually declines to a lower level. IgG-specific for SARS-CoV-2 N protein rapidly increases to a higher level after a secondary infection and retains in the body for several months even years.
|
| Alternative Names |
N; Nucleoprotein; N; Nucleocapsid protein; NC; Protein N |
| Abbreviation |
SARS-CoV-2 N Ab (IgG) |
| Uniprot No. |
P0DTC9 |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, plasma |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Infectious Diseases
|
| Assay Principle |
qualitative |
|
This assay mechanism is based on the qualitative enzyme immunoassay technique. The microtiter plate has been pre-coated with human SARS-CoV-2 nucleoprotein. Samples are pipetted into the wells with anti-human IgG conjugated HRP. Following a thorough wash, the TMB Substrate solution is added to the wells, eliciting a color change. The color develops in proportion to the amount of human SARS-CoV-2 N IgG antibody bound in the initial step. The addition of the stop solution to the wells terminates the color reaction. The color intensity can be measured at 450 nm via a microplate reader. The SARS-CoV-2 N IgG antibody titer in the samples is determined by referring to the negative control. It indicates the presence of SARS-CoV-2 N IgG antibody if the sample O.D. (optical density) is greater than or equal to the cutoff value (2.1×Average O.D. value of negative control). There is no SARS-CoV-2 N IgG antibody present in the sample if the O.D. is less than the cutoff value. |
| Precision |
| Intra-assay Precision (Precision within an assay): CV%<15% |
|
| Three samples of known concentration were tested twenty times on one plate to assess. |
| Inter-assay Precision (Precision between assays): CV%<15% |
|
| Three samples of known concentration were tested in twenty assays to assess. |
|
| Typical Data |
| Test parameter |
specification |
test result |
| Positive control |
≥0.6 |
0.867 |
| Negative control |
≤0.25 |
0.184 |
| Positive rate |
20，Positive |
100% |
| Negative rate |
20，Negative |
100% |
|
| Materials provided |
- A 96-well Coated assay plate --This microplate has been pre-coated with human SARS-CoV-2 nucleoprotein.
- Negative Control (1 x 800 μl) --It is free of the SARS-CoV-2 N IgG antibody and used to preclude the false positive.
- Positive Control (1 x 800 μl) --Used to evaluate the validity, stability, and comparability of the test results.
- HRP-conjugated anti-Human IgG antibody (100 x concentrate) (1 x 120 μl) --Act as the detection antibody.
- HRP-conjugate Diluent (1 x 20 ml) --Dilute the HRP-conjugated anti-Human IgG antibody.
- Sample Diluent (2 x 20 ml) --Dilute the sample solution.
- Wash Buffer (25 x concentrate) (1 x 20 ml) --Wash the unbound reagents.
- TMB Substrate (1 x 10 ml) --React with HRP, eliciting a chromogenic color reaction.
- Stop Solution (1 x 10 ml) --Stop the color reaction. The solution turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --Seal the microplate when incubation.
- An Instruction manual
|
| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator that can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer.
- Absorbent paper for blotting the microtiter plate.
- 50-300ul multi-channel micropipette
- 100ml and 500ml graduated cylinders.
- Deionized or distilled water.
- Pipette tips
- Timer
- Test tubes for dilution.
|
Troubleshooting
and FAQs |
ELISA kit FAQs |
| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
