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Human TP53(Tumor Protein p53) ELISA Kit
SKU: E-EL-H0910
Human TP53(Tumor Protein p53) ELISA Kit
| Detection Range | 78.13-5000 pg/mL |
| Sensitivity | 46.88 pg/mL |
Product Details
Properties
| Assay type | Sandwich-ELISA |
| Format | 96T |
| Assay time | 3.5h |
| Detection range | 78.13-5000 pg/mL |
| Sensitivity | 46.88 pg/mL |
| Sample type & Sample volume | serum, plasma and other biological fluids; 100μL |
| Sample size | Run up to 46 samples in duplicate and 30 samples in triplicate |
| Specificity | This kit recognizes Human TP53 in samples. No significant cross-reactivity or interference between Human TP53 and analogues was observed |
| Reproducibility | Both intra-CV and inter-CV are < 10% |
| Application | Quantitative determination of Human concentrations in human serum, plasma and other biological fluids |
Test Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human TP53. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TP53 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human TP53, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human TP53. You can calculate the concentration of Human TP53 in the samples by comparing the OD of the samples to the standard curve.
Kit Components and Storage
An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
|---|---|---|
| Micro ELISA Plate(Dismountable) | 96T: 8 wells ×12 strips 48T: 8 wells ×6 strips |
-20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial |
|
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
|
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
-20℃(Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 2-8℃(Protect from light) |
| Stop Solution | 1 vial, 10 mL | 2-8°C |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy |
Technical Data:
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and
high level Human TP53 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and
high level Human TP53 were tested on 3 different plates, 20 replicates in each plate,
respectively
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 264.04 | 720.64 | 2353.79 | 277.37 | 770.6 | 2533.22 |
| Standard deviation | 13.86 | 29.98 | 101.21 | 17.09 | 35.14 | 114.25 |
| CV (%) | 5.25 | 4.16 | 4.3 | 6.16 | 4.56 | 4.51 |
Recovery
The recovery of Human TP53 spiked at three different levels in samples throughout
the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 87-101 | 94 |
| EDTA plasma (n=8) | 85-98 | 91 |
| Cell culture media (n=8) | 93-104 | 99 |
Assay Procedures
 |
1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C |
 |
2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C |
 |
3. Aspirate and wash the plate for 3 times |
 |
4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times |
 |
5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C |
 |
6. Add 50μL Stop Solution |
 |
7. Read the plate at 450nm immediately. Calculation of the results |