CLIA Kit for Inositol Triphosphate (IP3)
InsP3; Inositol 1,4,5-Trisphosphate; Triphosphoinositol
- Product No.CCC037Ge
- Organism SpeciesPan-species (General) Same name, Different species.
- Test MethodCompetitive Inhibition
- Assay Length2h
- Detection Range123.5-10,000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 46.4pg/mL.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Download Instruction Manual
- UOM 48T96T 96T*5 96T*10 96T*100
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Specificity of the CLIA Kit for Inositol Triphosphate (IP3)
This assay has high sensitivity and excellent specificity for detection of Inositol Triphosphate (IP3).
No significant cross-reactivity or interference between Inositol Triphosphate (IP3) and analogues was observed.
Recovery of the CLIA Kit for Inositol Triphosphate (IP3)
Matrices listed below were spiked with certain level of Inositol Triphosphate (IP3) and the recovery rates were calculated by comparing the measured value to the expected amount of Inositol Triphosphate (IP3) in samples.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 97-104 | 101 |
EDTA plasma(n=5) | 89-96 | 92 |
heparin plasma(n=5) | 87-99 | 94 |
Precision of the CLIA Kit for Inositol Triphosphate (IP3)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Inositol Triphosphate (IP3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Inositol Triphosphate (IP3) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity of the CLIA Kit for Inositol Triphosphate (IP3)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Inositol Triphosphate (IP3) 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) | 79-95% | 82-97% | 94-101% | 93-105% |
EDTA plasma(n=5) | 88-101% | 84-104% | 80-95% | 85-96% |
heparin plasma(n=5) | 85-93% | 92-101% | 92-103% | 95-104% |
Stability of the CLIA Kit for Inositol Triphosphate (IP3)
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.
Assay procedure summary of the CLIA Kit for Inositol Triphosphate (IP3)
1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;
7. Read RLU value immediately.
Test principle of the CLIA Kit for Inositol Triphosphate (IP3)
The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Inositol Triphosphate (IP3). A competitive inhibition reaction is launched between biotin labeled Inositol Triphosphate (IP3) and unlabeled Inositol Triphosphate (IP3) (Standards or samples) with the pre-coated antibody specific to Inositol Triphosphate (IP3). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Inositol Triphosphate (IP3) in the sample. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is reverse proportional to the Inositol Triphosphate (IP3) level in the sample or standard.
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