- Product Name
- Hamster TNF-α(Tumor Necrosis Factor-α) ELISA Kit
- Alias
- TNF-α ELISA Kit, DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit
- Catalogue No.
- EHA0004
- Size
- 48T/96T
- Species
- Hamster
- UniProt No.
- A0A1U7RE37
- 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
- 3.125-200pg/ml
- Sensitivity
- 1.875pg/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with TNF-α , 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
- Assay Procedure Summary
-
- 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.
- 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 | 0.11 | 0.114 | 0.112 |
3.125 | 0.154 | 0.158 | 0.156 |
6.25 | 0.198 | 0.204 | 0.201 |
12.5 | 0.277 | 0.285 | 0.281 |
25 | 0.486 | 0.5 | 0.493 |
50 | 0.762 | 0.784 | 0.773 |
100 | 1.418 | 1.46 | 1.439 |
200 | 2.272 | 2.338 | 2.305 |
- Recovery
-
Add a certain amount of TNF-α into the sample. Calculate the recovery by comparing the measured value with the expected amount of TNF-α in the sample.
Sample Type | Recovery Range(%) | Average(%) |
serum(n=10) | 85-101 | 98 |
EDTA plasma(n=10) | 86-99 | 93 |
Heparin plasma(n=10) | 88-105 | 94 |
- Linearity
-
Dilute the sample with a certain amount of TNF-α at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type | 1:2 | 1:4 | 1:8 |
serum(n=10) | 80-104% | 84-100% | 88-102% |
EDTA plasma(n=10) | 85-104% | 86-102% | 84-98% |
Heparin plasma(n=10) | 90-98% | 80-96% | 81-98% |
- 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) | 6.31 | 25.76 | 98.89 | 6.13 | 26.15 | 98.43 |
Standard deviation | 0.31 | 1.03 | 3.99 | 0.25 | 1.12 | 4.28 |
CV(%) | 4.88 | 3.98 | 4.03 | 4.06 | 4.28 | 4.35 |
- 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 |