SARS-CoV-2 Neutralizing Antibody ELISA Kit

Instructions
Code CSB-EL33243HU
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

CUSABIO's SARS-CoV-2 Neutralizing Antibody ELISA Kit is intended for qualitative measurement of neutralizing antibodies in human serum or plasma. SARS-CoV-2 neutralizing antibodies directly target the SARS-CoV-2 S receptor-binding domain (RBD), blocking the protein's interaction with the host ACE2 receptor, thus interfering with the viral attachment to the cell as well as its entry into the cell. When using the reference neutralizing antibody provided in the kit, the assay has a detection range of 9.75-10000 ng/ml, a sensitivity of as low as 9.75 ng/ml, and Intra- /Inter-assay Precision low than 15%. This detection kit is simple in operation and scalable in sample size (high throughput). Multiple validations display it has excellent sensitivity and high specificity. It is conducive to identify individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection. And it should not be used for the diagnosis of acute SARS-CoV-2 infection.

Alternative Names 2019 Novel Coronavirus; Coronavirus; CoV; COVID-19 virus; HCoV-2; Human Coronavirus 2019; SARS2; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2
Abbreviation SARS-CoV-2 neutralizing Ab
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
Measurement Competitive

The assay employs the competitive inhibition enzyme immunoassay technique as well as a colorimetric reaction, in which the SARS-CoV-2 neutralizing antibodies in the sample and standard compete with the HRP-conjugated ACE2 to bind to the pre-coated SARS-CoV-2 S RBD. After the sequential addition of the TMB Substrate Solution and the Stop solution, the solution color develops in opposite to the amount of SARS-CoV-2 neutralizing antibodies in the sample. The presence of neutralizing antibodies in the sample can be detected by a reduction signal of HRP-ACE2/SARS-CoV-2 S RBD binding.
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
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

ng/ml OD1 OD2 Average
10000 0.099 0.092 0.096
5000 0.145 0.136 0.141
2500 0.194 0.183 0.189
1250 0.274 0.263 0.269
625 0.344 0.371 0.358
312 0.519 0.528 0.524
156 0.901 0.907 0.904
78 1.509 1.412 1.461
39 1.787 1.723 1.755
19.5 2.121 1.958 2.040
9.75 2.233 2.138 2.186
0 2.394 2.328 2.361
Materials provided
    • One 96-well Microtiter plate ( coated Microwells) -- This microwell plate has been pre-coated with recombinant SARS-CoV-2 S RBD antigen and can be dismountable into 12 strips of 8 wells.
    • A bottle of Standard (10 x concentrate; 200 μl/vial) -- Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
    • A bottle of HRP-conjugated ACE2 (100 x concentrate; 60 μl/vial) -- ACE2 interacts with SARS-CoV-2 S RBD, mimicking the process of SARS-CoV-2 entry into the host cells. HRP catalyzes the TMB Substrate solution, eliciting a chromogenic reaction.
    • A bottle of HRP-conjugate Diluent (10 ml/vial) -- Dilute the HRP-conjugated ACE2 solution.
    • Two bottles of Sample Diluent (20 ml/vial) -- Reconstitute the standard and dilute the sample to an appropriate concentration.
    • A bottle of Wash Buffer (25 x concentrate; 20 ml/vial) -- Wash away unbound or free substances.
    • A bottle of TMB Substrate (10 ml/vial) -- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
    • A bottle of Stop Solution (10 ml/vial) -- Stop the color reaction. The solution color immediately turns from blue to yellow.
    • Four Adhesive Strips (For 96 wells) -- Seal the plate when incubation.
    • One 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

Citations

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 Q&A
Q:

How Do SARS-CoV-2 Neutralizing Antibodies Work?

A:

Neutralizing antibodies (NAbs) are a type of special antibodies with the capacity of neutralizing viral toxicity. The invasion of SARS-CoV-2 into host cells depends on the interaction between the viral S protein and the ACE2 receptor on host cells. NAbs against SARS-CoV-2 specifically bind to the RBD within the S protein of SARS-CoV-2, blocking RBD-ACE2 receptor interaction, thus preventing the virus infect the host cells and clearing the virus. For more details about Neutralizing antibodies, please read this article: https://www.cusabio.com/c-21014.html

Q:

Why should I choose the SARS-CoV-2 neutralizing antibody ELISA kit from CUSABIO?

A:
1. Simple, convenient and fast;
2. Operation can be completed in any laboratory setting;
3. Strong specificity, small inter-assay variation.
Q:

What is the principle of the SARS-CoV-2 Neutralizing Antibody ELISA Kit?

A:
This assay is based on competitive method. The kit detects circulating neutralizing antibodies against SARS-CoV-2 that block the interaction between the receptor binding domain (RBD) of the viral spike glycoprotein with the ACE2 cell surface receptor.
Q:

Which applications can I use the SARS-CoV-2 neutralizing antibody ELISA kit for?

A:
1. Viral spike glycoprotein (RBD)/ACE2 receptor interaction studies;
2. Vaccine and Biologics development;
3. COVID-19 Epidemiology Studies.
Q:

What is the assay procedure?

A:

1. Prepare all reagents, samples, and working standards as instructed.
2. Refer to the Assay Layout Sheet to calculate the number of the wells (including a blank well) to be used and put the remaining wells and the desiccant back into the pouch to seal and store at 4 °C.
3. Add 50 µl of standard or sample (may require dilution) to each well, followed by 50 µl of HRP-conjugated ACE2 (not to the blank well). Shake the plate gently for 1 minute. Seal and incubate for 1 hour at  37 °C.
4. After thorough washing, add 90 μl of TMB Substrate Solution to each well. Incubate for 20 minutes at 37°C.
5. Add 50 µl Stop Solution to each well and gently tap the plate to ensure thorough mixing.
6. Read the OD (optical density) value at 450 nm within 5 minutes using a microplate reader.

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