ELISA Kit for Myoglobin (MYO)
PVALB; MB
| Organism species | Homo sapiens (Human) |
| Product No. | SEA480Hu |
| Sample type | Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. |
| Format | 96-well strip plate |
| Assay length | 4.5 hours |
| Detection range | 0.781-50ng/mL The standard curve concentrations used for the ELISA’s were 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.125ng/mL, 1.563ng/mL, 0.781ng/mL |
| Sensitivity | The minimum detectable dose of this kit is typically less than 0.30ng/mL. |
Specificity
This assay has high sensitivity and excellent specificity for detection of Myoglobin (MYO).
No significant cross-reactivity or interference between Myoglobin (MYO) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Myoglobin (MYO) and the recovery rates were calculated by comparing the measured value to the expected amount of Myoglobin (MYO) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 93-101 | 96 |
| EDTA plasma(n=5) | 95-102 | 98 |
| heparin plasma(n=5) | 78-90 | 81 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Myoglobin (MYO) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Myoglobin (MYO) 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 Myoglobin (MYO) 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) | 89-104% | 90-101% | 93-102% | 84-101% |
| EDTA plasma(n=5) | 79-105% | 82-102% | 86-99% | 88-103% |
| heparin plasma(n=5) | 86-93% | 84-99% | 93-105% | 86-98% |
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.
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 |
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 Myoglobin (MYO). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Myoglobin (MYO). 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 Myoglobin (MYO), 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 Myoglobin (MYO) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
