This kit is based on indirect ELISA detection method and takes 3h assay time. The microplate provided in this kit has been precoated with antigen. HRP-antibody is used as detection antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add HRP-detection antibody, then it binds with anti- SARS-CoV-2 (S protein) IgG bound to precoated antibody. Wash unbound components 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 anti- SARS-CoV-2 (S protein) IgG in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Manual- Product Name
- Hamster anti- SARS-CoV-2 (S protein) IgG ELISA Kit
- Alias
- SARS-CoV-2(S protein) IgG ELISA Kit
- Catalogue No.
- EHA0025
- Size
- 48T/96T
- Species
- Hamster
- Sample Type
- Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
- Detection Method
- Indirect ELISA Sandwich
- Detection Wavelength
- OD450
- Reaction Duration
- 3 hours
- Range
- 1.563-100ng/ml
- Sensitivity
- 0.938ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with anti- SARS-CoV-2 (S protein) IgG , 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
-
- Step 1: Wash the plate twice before adding the standard and sample.
- Step 2: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
- Washing: Wash the plate three times and immerse for 1min each time.
- Step 3: Add 100ul HRP-antibody 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 0.122 0.126 0.124 1.562 0.257 0.265 0.261 3.125 0.417 0.429 0.423 6.25 0.773 0.795 0.784 12.5 1.303 1.341 1.322 25 1.668 1.716 1.692 50 2.098 2.158 2.128 100 2.245 2.311 2.278 - Recovery
-
Add a certain amount of anti- SARS-CoV-2 (S protein) IgG into the sample. Calculate the recovery by comparing the measured value with the expected amount of anti- SARS-CoV-2 (S protein) IgG in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 86-96 90 EDTA plasma(n=10) 90-104 97 Heparin plasma(n=10) 87-103 96 - Linearity
-
Dilute the sample with a certain amount of anti- SARS-CoV-2 (S protein) IgG at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 86-104% 82-101% 80-94% EDTA plasma(n=10) 90-101% 92-100% 90-96% Heparin plasma(n=10) 88-100% 84-96% 84-97% - 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) 2.91 11.37 47.35 3.17 11.79 48.66 Standard deviation 0.15 0.56 2.28 0.16 0.73 2.31 CV(%) 5.22 4.89 4.82 4.95 6.2 4.75 - 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