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| 货号： | SEA553Bo |
| 样本： | Serum, plasma, tissue homogenates, cell lysates and other biological fluids. |
| 应用： | 体外科学研究 |
| 检测方法： | 酶联免疫吸附试验法 |
| 检测限： | 31.25-2000pg/mL |
| 供应商： | 研卉生物 |
| 数量： | 1-2周 |
| 规格： | 96T |
基质金属蛋白酶9(MMP9)检测试剂盒(酶联免疫吸附试验法)
ELISA Kit for Matrix Metalloproteinase 9 (MMP9)
ELISA Kit for Matrix Metalloproteinase 9 (MMP9)
ELISA Kit for Matrix Metalloproteinase 9 (MMP9)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
| Organism species | Bos taurus; Bovine (Cattle) |
| Product No. | SEA553Bo |
| Sample type | Serum, plasma, tissue homogenates, cell lysates and other biological fluids. |
| Format | 96-well strip plate |
| Assay length | 4.5 hours |
| Detection range | 31.25-2000pg/mL The standard curve concentrations used for the ELISA’s were 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL |
| Sensitivity | The minimum detectable dose of this kit is typically less than 11.3pg/mL. |
Specificity
This assay has high sensitivity and excellent specificity for detection of Matrix Metalloproteinase 9 (MMP9).
No significant cross-reactivity or interference between Matrix Metalloproteinase 9 (MMP9) and analogues was observed.
No significant cross-reactivity or interference between Matrix Metalloproteinase 9 (MMP9) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Matrix Metalloproteinase 9 (MMP9) and the recovery rates were calculated by comparing the measured value to the expected amount of Matrix Metalloproteinase 9 (MMP9) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 78-101 | 84 |
| heparin plasma(n=5) | 94-101 | 98 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Metalloproteinase 9 (MMP9) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Matrix Metalloproteinase 9 (MMP9) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Matrix Metalloproteinase 9 (MMP9) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Matrix Metalloproteinase 9 (MMP9) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 94-103% | 78-88% | 79-95% | 96-105% |
| heparin plasma(n=5) | 83-97% | 79-95% | 92-99% | 82-89% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
| Reagents | Quantity | Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 | Plate sealer for 96 wells | 4 |
| Standard | 2 | Standard Diluent | 1×20mL |
| Detection Reagent A | 1×120µL | Assay Diluent A | 1×12mL |
| Detection Reagent B | 1×120µL | Assay Diluent B | 1×12mL |
| TMB Substrate | 1×9mL | Stop Solution | 1×6mL |
| Wash Buffer (30 × concentrate) | 1×20mL | Instruction manual | 1 |
Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
8. Add 50µL Stop Solution. Read at 450nm immediately.
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
8. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Matrix Metalloproteinase 9 (MMP9). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Matrix Metalloproteinase 9 (MMP9). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Matrix Metalloproteinase 9 (MMP9), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Matrix Metalloproteinase 9 (MMP9) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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