COVID-19 Neutralizing Antibody ELISA Kit

Category:

This kit is based on Double antigen-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with antigen. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antigen. Then, it binds with COVID-19 Neutralizing Antibody bound to precoated antigen. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of COVID-19 Neutralizing Antibody in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: 
EU3137
Alias: 
COVID-19 Neutralizing Antibody ELISA Kit
Species: 
Universal
Range: 
1.563-100ng/ml
Sensitivity: 
0.938ng/ml
  • SPECIFICATIONS
Product Name
COVID-19 Neutralizing Antibody ELISA Kit
Alias
COVID-19 Neutralizing Antibody ELISA Kit
Catalogue No.
EU3137
Size
48T/96T
Species
Universal
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
Sandwich ELISA, Double Antigen
Detection Wavelength
OD450
Reaction Duration
4 hours
Range
1.563-100ng/ml
Sensitivity
0.938ng/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with COVID-19 Neutralizing Antibody , no obvious cross reaction with other analogues.
Required Instruments and Reagents
  1. Microplate reader (wavelength: 450nm)
  2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
  3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  5. Sterile tubes and Eppendorf tubes with disposable tips
  6. Absorbent paper and loading slot
  7. Deionized or distilled water
Assay Procedure Summary
Elisa实验原理图
  • Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
  • Washing: Wash the plate twice without immersing.
  • Step 2: Add 100ul biotin-antigen working solution, seal the plate and statically incubate for 60 minutes at 37°C.
  • Washing: Wash the plate three times and immerse for 1min each time.
  • Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
  • Washing: Wash the plate five times and immerse for 1min each time.
  • Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
  • Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.
Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(ng/ml) OD-1 OD-2 Average
0 0.086 0.087 0.087
1.562 0.231 0.233 0.232
3.125 0.285 0.272 0.279
6.25 0.365 0.348 0.357
12.5 0.512 0.484 0.498
25 0.952 0.889 0.921
50 1.581 1.422 1.502
100 2.331 2.426 2.379
EU3137 Standard Curve Image
Recovery

Add a certain amount of COVID-19 Neutralizing Antibody into the sample. Calculate the recovery by comparing the measured value with the expected amount of COVID-19 Neutralizing Antibody in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 87-104 94
EDTA plasma(n=10) 86-105 99
Heparin plasma(n=10) 86-105 92
Linearity

Dilute the sample with a certain amount of COVID-19 Neutralizing Antibody at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 86-97% 86-98% 80-93%
EDTA plasma(n=10) 87-105% 85-100% 81-97%
Heparin plasma(n=10) 85-103% 86-100% 81-100%
Precision(%)

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/ml) 3.02 12.5 48.48 3.04 12.92 49.79
Standard deviation 0.15 0.62 2.48 0.16 0.63 2.48
CV(%) 4.82 4.98 5.12 5.23 4.88 4.98
Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100