ELISA Kit for Sphingosine-1-Phosphate (S1P)

FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
    • ELISA Kit for Sphingosine-1-Phosphate (S1P)Packages (Simulation)
    • ELISA Kit for Sphingosine-1-Phosphate (S1P)Packages (Simulation)
    • ELISA Kit for Sphingosine-1-Phosphate (S1P)Results demonstration
    • CEG031Ge.jpgTypical Standard Curve
    • CertificateISO9001: 2008, ISO13485: 2003 Registered

    Specificity

    This assay has high sensitivity and excellent specificity for detection of Sphingosine-1-Phosphate (S1P).
    No significant cross-reactivity or interference between Sphingosine-1-Phosphate (S1P) and analogues was observed.

    Recovery

    Matrices listed below were spiked with certain level of recombinant Sphingosine-1-Phosphate (S1P) and the recovery rates were calculated by comparing the measured value to the expected amount of Sphingosine-1-Phosphate (S1P) in samples.

    MatrixRecovery range (%)Average(%)
    serum(n=5)94-10598
    EDTA plasma(n=5)81-9886
    heparin plasma(n=5)80-10382

    Precision

    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sphingosine-1-Phosphate (S1P) were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Sphingosine-1-Phosphate (S1P) 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 Sphingosine-1-Phosphate (S1P) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

    Sample1:21:41:81:16
    serum(n=5)89-101%94-103%82-99%82-104%
    EDTA plasma(n=5)81-94%90-98%78-96%84-97%
    heparin plasma(n=5)98-105%78-105%86-95%78-96%

    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

    ReagentsQuantityReagentsQuantity
    Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
    Standard2Standard Diluent1×20mL
    Detection Reagent A1×120µLAssay Diluent A1×12mL
    Detection Reagent B1×120µLAssay Diluent B1×12mL
    TMB Substrate1×9mLStop Solution1×6mL
    Wash Buffer (30 × concentrate)1×20mLInstruction manual1

    Assay procedure summary

    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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
    7. Add 50µL Stop Solution. Read at 450 nm immediately.

    ELISA Kit for Sphingosine-1-Phosphate (S1P)

    Test principle

    This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Sphingosine-1-Phosphate (S1P) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Sphingosine-1-Phosphate (S1P) and unlabeled Sphingosine-1-Phosphate (S1P) (Standards or samples) with the pre-coated antibody specific to Sphingosine-1-Phosphate (S1P). 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 Sphingosine-1-Phosphate (S1P) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Sphingosine-1-Phosphate (S1P) in the sample.

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