| Product Name | Mouse IFN-γ(Interferon Gamma) ELISA Kit |
| UniProt ID | P01580 |
| Sample Type | Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples |
| Detection Method | Sandwich ELISA, Double Antibody |
| Detection Wavelength | OD450 |
| Reaction Duration | 4 hours |
| Range | 31.25-2000pg/ml (1ng=8430mIU) |
| Storage | 2-8°C(Sealed), Don't cryopreserve |
| Specificity | Specifically binds with IFN-γ , no obvious cross reaction with other analogues. |
ELISA Kit Components
| Kit Components | Item | Size(48T) | Size(96T) | Storage Condition for Opened Kit |
| E001 | ELISA Microplate(Dismountable) | 8×6 | 8×12 | Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C |
| E002 | Lyophilized Standard | 1vial | 2vial | Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C |
| E003 | Biotin-labeled Antibody(Concentrated, 100X) | 60ul | 120ul | 2-8°C (Avoid Direct Light) |
| E034 | HRP-Streptavidin Conjugate(SABC, 100X) | 60ul | 120ul | |
| E024 | TMB Substrate | 5ml | 10ml | |
| E039 | Sample Dilution Buffer | 10ml | 20ml | 2-8°C |
| E040 | Antibody Dilution Buffer | 5ml | 10ml | |
| E049 | SABC Dilution Buffer | 5ml | 10ml | |
| E026 | Stop Solution | 5ml | 10ml | |
| E038 | Wash Buffer(Concentrated, 25X) | 15ml | 30ml | |
| E006 | Plate Sealer | 3 pieces | 5 pieces | |
| E007 | Product Description | 1 copy | 1 copy |
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
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.(pg/ml) | OD-1 | OD-2 | Average |
| 0 | 0.059 | 0.062 | 0.06 |
| 31.25 | 0.104 | 0.108 | 0.106 |
| 62.5 | 0.149 | 0.155 | 0.152 |
| 125 | 0.217 | 0.226 | 0.222 |
| 250 | 0.382 | 0.398 | 0.39 |
| 500 | 0.646 | 0.673 | 0.66 |
| 1000 | 1.299 | 1.352 | 1.325 |
| 2000 | 2.297 | 2.391 | 2.344 |
Recovery
Add a certain amount of IFN-γ into the sample. Calculate the recovery by comparing the measured value with the expected amount of IFN-γ in the sample.
| Sample Type | Recovery Range(%) | Average(%) |
| serum(n=10) | 91-100 | 97 |
| EDTA plasma(n=10) | 87-105 | 96 |
| Heparin plasma(n=10) | 85-102 | 92 |
Linearity
Dilute the sample with a certain amount of IFN-γ at 1:2, 1:4 and 1:8 to get the recovery range.
| Sample Type | 1:2 | 1:4 | 1:8 |
| serum(n=10) | 89-99% | 89-99% | 84-100% |
| EDTA plasma(n=10) | 91-100% | 83-92% | 80-93% |
| Heparin plasma(n=10) | 85-105% | 82-99% | 82-87% |
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 (pg/ml) | 62.47 | 235.1 | 1003.7 | 60.14 | 240.8 | 988.9 |
| Standard deviation | 3.52 | 12.06 | 51.99 | 3.06 | 12.09 | 53.2 |
| CV(%) | 5.63 | 5.13 | 5.18 | 5.09 | 5.02 | 5.38 |
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 |
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