This kit is based on indirect ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with antigen. Biotinylated antibody is used as detection antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with COVID-19 Spike protein (S2) IgA 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 Spike protein (S2) IgA in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Manual- Product Name
- Human COVID-19 Spike protein (S2) IgA ELISA Kit
- Alias
- COVID-19 Spike protein (S2) IgA ELISA Kit
- Catalogue No.
- EH5017
- Size
- 48T/96T
- Species
- Human
- Sample Type
- Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
- Detection Method
- Indirect ELISA
- Detection Wavelength
- OD450
- Reaction Duration
- 2 hours
- Range
- 0.391-25ng/ml
- Sensitivity
- 0.234ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with COVID-19 Spike protein (S2) IgA , no obvious cross reaction with other analogues.
- Required Instruments and Reagents
-
- Microplate reader (wavelength: 450nm)
- 37°C incubator (CO2 incubator for cell culture is not recommenced.)
- Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
- Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
- Sterile tubes and Eppendorf tubes with disposable tips
- Absorbent paper and loading slot
- Deionized or distilled water
- Assay Procedure Summary
-
- 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 0.091 0.088 0.391 0.172 0.181 0.176 0.781 0.241 0.253 0.246 1.562 0.374 0.393 0.382 3.125 0.653 0.686 0.666 6.25 1.046 1.099 1.067 12.5 1.602 1.684 1.635 25 2.15 2.26 2.194 - Recovery
-
Add a certain amount of COVID-19 Spike protein (S2) IgA into the sample. Calculate the recovery by comparing the measured value with the expected amount of COVID-19 Spike protein (S2) IgA in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 85-101 95 EDTA plasma(n=10) 86-100 91 Heparin plasma(n=10) 86-104 95 - Linearity
-
Dilute the sample with a certain amount of COVID-19 Spike protein (S2) IgA at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 87-101% 83-101% 86-92% EDTA plasma(n=10) 85-104% 85-96% 80-86% Heparin plasma(n=10) 94-104% 82-96% 82-92% - 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) 0.84 3.02 12.55 0.82 3.24 13.38 Standard deviation 0.04 0.15 0.4 0.04 0.17 0.69 CV(%) 5.21 5.08 3.22 4.69 5.12 5.12 - 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