ELISA Kit for Calprotectin (CALPRO)

MRP-8/MRP-14; S100A8/A9

FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
    • ELISA Kit for Calprotectin (CALPRO)Packages (Simulation)
    • ELISA Kit for Calprotectin (CALPRO)Packages (Simulation)
    • ELISA Kit for Calprotectin (CALPRO)Results demonstration
    • SEK504Mu.jpgTypical Standard Curve
    • CertificateISO9001: 2008, ISO13485: 2003 Registered

    Specificity

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

    Recovery

    Matrices listed below were spiked with certain level of recombinant Calprotectin (CALPRO) and the recovery rates were calculated by comparing the measured value to the expected amount of Calprotectin (CALPRO) in samples.

    MatrixRecovery range (%)Average(%)
    serum(n=5)78-10590
    EDTA plasma(n=5)96-10399
    heparin plasma(n=5)86-9490

    Precision

    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Calprotectin (CALPRO) were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Calprotectin (CALPRO) 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 Calprotectin (CALPRO) 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)82-91%96-104%95-102%80-101%
    EDTA plasma(n=5)87-97%86-95%83-93%92-101%
    heparin plasma(n=5)86-101%91-102%78-94%81-96%

    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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
    3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
    4. Aspirate and wash 3 times;
    5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
    6. Aspirate and wash 5 times;
    7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
    8. Add 50µL Stop Solution. Read at 450nm immediately.

    ELISA Kit for Calprotectin (CALPRO)

    Test principle

    The microplate provided in this kit has been pre-coated with an antibody specific to S100A8. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to S100A9. 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 calprotectin, 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 calprotectin in the samples is then determined by comparing the O.D. of the samples to the standard curve.

    Related products

    Catalog No.Organism species: Mus musculus (Mouse)Applications
    RPK504Mu01Recombinant Calprotectin (CALPRO)Positive Control; Immunogen; SDS-PAGE; WB.
    SEK504MuELISA Kit for Calprotectin (CALPRO)Enzyme-linked immunosorbent assay for Antigen Detection.
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