Description
| Product Name: | Human TNF-alpha ELISA Kit (High Sensitivity) |
| Alias: | TNFα ELISA Kit, Tumor Necrosis Factor Alpha ELISA Kit, TNF-α ELISA Kit, DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit |
| SKU: | AEFI01066 |
| Size: | 96T |
| Species: | Human |
| UniProt No: | P01375 |
| 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: | 3.12-200pg/ml |
| Sensitivity: | 1.875pg/ml |
| Storage: | 2-8°C(Sealed), Don't cryopreserve. |
| Specificity: | Specifically binds with TNFα, no obvious cross reaction with other analogues. |
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C |
| Lyophilized Standard | 2 vials | Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C |
| Biotin-labeled Antibody (Concentrated, 100X) | 120 ul | 2-8°C (Avoid Direct Light) |
| HRP-Streptavidin Conjugate (SABC, 100X) | 120 ul | |
| TMB Substrate | 10 ml | |
| Sample Dilution Buffer | 20 ml | 2-8°C |
| Antibody Dilution Buffer | 10 ml | |
| SABC Dilution Buffer | 10 ml | |
| Stop Solution | 10 ml | |
| Wash Buffer (Concentrated, 25X) | 30 ml | |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy |
Other materials and equipment required:
- Microplate reader (wavelength: 450nm)
- 37°C incubator (CO2 incubator for cell culture is not recommended.)
- 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
| Recovery: | Add a certain amount of TNFα into the sample. Calculate the recovery by comparing the measured value with the expected amount of TNFα in the sample. | ||||||||||||||||
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| Linearity: | Dilute the sample with a certain amount of TNFα at 1:2, 1:4 and 1:8 to get the recovery range. | ||||||||||||||||
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| Stability: | Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data. | ||||||||||||||||
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| Precision(%): | Intra-Assay: CV<8% Inter-Assay: CV<10% |
| 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-antibody 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. |
