Description
Specifications
This assay has high sensitivity and excellent specificity for the detection of Interleukin 10 (IL10).
No significant cross-reactivity or interference between Interleukin 10 (IL10) and analogs was observed.
Recovery
Matrices listed below were spiked with a certain level of recombinant Interleukin 10 (IL10) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 10 (IL10) in samples.
Matrix | Recovery range (%) | Average(%) |
---|---|---|
serum(n=5) | 91-105 | 97 |
EDTA plasma(n=5) | 78-99 | 95 |
heparin plasma(n=5) | 94-105 | 98 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level Interleukin 10 (IL10) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle, and high-level Interleukin 10 (IL10) were tested on 3 different plates, with 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 Interleukin 10 (IL10) 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) | 78-99% | 89-96% | 85-98% | 92-102% |
EDTA plasma(n=5) | 86-102% | 93-101% | 95-103% | 99-105% |
heparin plasma(n=5) | 80-101% | 78-98% | 78-96% | 95-105% |
Stability
The stability of the 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 conditions.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, and 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 |
Procedure
1. Prepare all reagents, samples, and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate for 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate for 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate for 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.