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-MPV 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-MPV IgG in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Manual- Product Name
- Human Anti-MPV IgG(Anti-Monkeypox Virus IgG) ELISA Kit
- Alias
- MPXV IgG ELISA Kit, Monkeypox virus IgG ELISA Kit
- Catalogue No.
- EH4996
- 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
- 1.5 hours
- Range
- 3.125-200ng/ml
- Sensitivity
- 1.875ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with Anti-MPV 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
-
- 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 0.045 0.047 0.046 3.125 0.139 0.146 0.142 6.25 0.214 0.225 0.218 12.5 0.355 0.373 0.362 25 0.623 0.655 0.636 50 0.996 1.046 1.016 100 1.57 1.65 1.602 200 2 2.102 2.041 - Recovery
-
Add a certain amount of Anti-MPV IgG into the sample. Calculate the recovery by comparing the measured value with the expected amount of Anti-MPV IgG in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 87-105 96 EDTA plasma(n=10) 90-97 94 Heparin plasma(n=10) 91-104 97 - Linearity
-
Dilute the sample with a certain amount of Anti-MPV 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) 92-102% 88-103% 91-102% EDTA plasma(n=10) 84-90% 82-100% 82-91% Heparin plasma(n=10) 87-100% 80-97% 83-99% - 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) 5.8 23.4 101.5 5.98 23.66 97.9 Standard deviation 0.3 1.19 5.51 0.31 1.28 6.29 CV(%) 5.22 5.08 5.43 5.12 5.43 6.43 - 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