| ID | Components | Size |
| TAS022-C01 | Pre-coated with Human ACE2 Protein Microplate | 1 plate |
| TAS022-C02 | Negative Control | 100 μL |
| TAS022-C03 | HRP-SARS-CoV-2 Spike RBD | 10 μg |
| TAS022-C04 | 10xWashing Buffer | 50 mL |
| TAS022-C05 | Dilution Buffer | 50 mL |
| TAS022-C06 | Substrate Solution | 12 mL |
| TAS022-C07 | Stop Solution | 7 mL |
| TAS022-C08 | Positive Control | 100 μL |
Background/背景
The newly identified Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) poses continual threat to human health due to rapid transmission worldwide. The Anti SARS-CoV-2 Neutralizing Antibody Titer Serologic Assay Kit to test the protective neutralizing antibody levels in post-vaccination serum or convalescent serum.
Application/应用
This assay kit is used to measure the levels of Anti-SARS-CoV-2 neutralizing antibody which interfere ACE2/S protein RBD interaction.
It is for research use only.
Reconstitution/重构方法
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
Storage/存储
The unopened kit is stable for at least 1 year from the date of manufacture if stored at 2°C to 8°C, and the opened kit is stable for up to 1 month from the date of opening at 2°C to 8°C.
It is recommended not to freeze thaw more than 3 times for lyophilized proteins.
Assay Principles/原理
The Anti-SARS-CoV-2 Neutralizing Antibody Titer Serologic Assay kit is a competitive ELISA. Immobilized Human ACE2 protein on the microplate are incubated with samples and Spike RBD-HRP. Presence of anti-SARS-CoV-2 neutralizing antibody in the samples competes with immobilized Human ACE2 protein to bind Spike RBD-HRP. The intensity of assay signal decreases proportionally to the presence of Anti-SARS-CoV-2 neutralizing antibody.
The kit analysis includes 5 simple steps:
a) All reagents are warmed to room temperature (20℃-25℃) before use.
b) Series of sample and control antibody dilution are prepared with dilution buffer. Solution of Spike RBD-HRP is prepared with dilution buffer.
c) The sample dilution is added to plate followed by Spike RBD-HRP solution. For semiquantitative analysis, dilution of control neutralizing antibody is add to the plate followed by Spike RBD-HRP solution.
d) After incubation, plates are washed and TMB or other colorimetric HRP substrate is addded.
e) The substrate reaction is stopped by add diluted acid. Absorbance (OD) is calculated as the absorbance at 450 nm minus the absorbance at 650 nm to remove background noise prior to statistical analysis. The OD Value reflects the amount of protein bound.
Typical Data/典型数据
Inhibition of Human ACE2: SARS-CoV-2 Spike RBD (HRP) binding by Anti-SARS-CoV-2 Neutralizing Antibody measured by competitive ELISA Assay.
Serial dilutions of Anti-SARS-CoV-2 Neutralizing Antibody (Cat. No. TAS022-C02) (1:2 serial dilutions, from 1250 ng/mL to 1.2 ng/mL) was added into Human ACE2: SARS-CoV-2 Spike RBD (HRP) binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points prior to log transformation and curve fitting. A representative result showed that the IC50 was calculated to be 60 ng/mL (QC tested).
Protocol
