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Cortisol ELISA Kit

Reaktivität: Hormone Colorimetric Competition ELISA 0-500 ng/mL
Produktnummer ABIN996929
  • Target Alle Cortisol ELISA Kits anzeigen
    Cortisol
    Reaktivität
    • 8
    • 5
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Hormone
    Nachweismethode
    Colorimetric
    Methodentyp
    Competition ELISA
    Detektionsbereich
    0-500 ng/mL
    Untere Nachweisgrenze
    0 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    The Quantitative Determination of Total Cortisol Concentration in Human Serum or Plasma by a Microplate Enzyme Immunoassay.
    Analytische Methode
    Quantitative
    Sensitivität
    0.37 μg/dL
    Benötigtes Material
    1. Pipette capable of delivering 25μL, 50μL and 100μL volumes with a precision of better than 1.5 %.
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  • Kommentare

    Quality Control:
    Each laboratory should assay controls at levels in the low, normal and high range for monitoring assay performance. These controls should be treated as unknowns and values determined in every test procedure performed. Quality control charts should be maintained - follow the performance of the supplied reagents. Pertinent statistical methods should be employed - ascertain trends. The individual laboratory should set acceptable assay performance limits. In addition, maximum absorbance should be consistent with past experience. Significant deviation from established performance can indicate unnoticed change in experimental conditions or degradation of kit reagents. Fresh reagents should be used - determine the reason for the variations.

    Plattentyp
    Pre-coated
    Aufbereitung der Proben

    The specimens shall be blood, serum or plasma in type and the usual precautions in the collection of venipuncture samples should be observed. The accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain redtop venipuncture tube without additives Ci 2 3 9 10. 11. 12. 13.

    Testdurchführung

    Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20-27 °C). T est Procedure should be performed by a skilled individual or trained professional**
    Note: Dilute the samples suspected of concentrations higher than 50 μg/dL 1:5 and 1:10 with cortiso '0' μg/dL patient serum.

    Ergebnisberechnung

    A dose response curve is used to ascertain the concentration of cortisol in unknown specimens.
    1. Record the absorbance obtained from the printout of the microplate reader as outlined in Example
    1.

    2. Plot the absorbance for each duplicate serum reference versus the corresponding cortisol concentration in μg/dL on linear graph paper (do not average the duplicates of the serum references before plotting).

    3. Connect the points with a best -fit curve. To determine the concentration of cortisol for an unknown, locate the average absorbance of the duplicates for each unknown on the vertical axis of the graph, find the intersecting point on the curve, and read the concentration (in mg/dL ) from the horizontal axis of the graph (the duplicates of the unknown may be averaged as indicated). In the following example, the average absorbance (
    1.071) intersects the dose response curve at (10.2 ng/dL) cortisol concentration (See figure 1). Note: Computer data reduction software designed for ELISA assays may also be used for the data reduction. If such software should be ascertained. 4 5 8 9 10. 1
    1. Sample I.D. Well Number Abs (A) Mean Abs (B) Value Cal A A1

    2.483

    2.543 0 B1

    2.575 Cal B C1

    2.150

    2.194
    1.0 D1

    2.186 Cal C E1
    1.573
    1.585
    4.0 F1
    1.597 Cal D G1
    1.103
    1.084 10 H1
    1.065 Cal E A2 0.726 0.725 20 B2 0.724 Cal F C2 0.347 0.350 50 D2 0.353 Ctrl 1 E2
    1.624
    1.617

    3.74 F2
    1.611 Ctrl 2 G2 0.770 0.760 18.57 H2 0.749 Patient 1 A3
    1.056
    1.071 10.24 B3
    1.086 *The data presented in Example 1 is for illustration only and should not be used in lieu of a standard curve prepared with each assay. Q.C. PARAMETERS In order for the assay results to be considered valid the following criteria should be met:
    1. The absorbance (OD) of calibrator 0 mg/dL should be >
    1.

    3.

    2. Four out of six quality control pools should be within the established ranges. RISK ANALYSIS A. Assay Performance
    1. It is important that the time of reaction in each well is held constant for reproducible results.

    2. Pipetting of samples should not extend beyond ten (10) minutes to avoid assay drift.

    3. Highly lipemic, hemolyzed or grossly contaminated specimen(s) should not be used. Ci If more than one (1) plate is used, it is recommended to repeat the dose response curve. The addition of the substrate solution initiates a kinetic reaction, which is terminated by the addition of the stop solution. Therefore, the substrate and stop solution should be added in the same sequence to eliminate any time-deviation during reaction. Plate readers measure vertically. Do not touch the bottom of the wells. Failure to remove adhering solution adequately in the aspiration or decantation wash step(s) may result in poor replication and spurious results. Use components from the same lot. No intermixing of reagents from different batches. Accurate and precise pipetting, as well as following the exact time and temperature requirements prescribed are essential. Any deviation from DAI IFU yield inaccurate results. All applicable national standards, regulations and laws, including, but not limited to, good laboratory procedures, must be strictly followed to ensure compliance and proper device usage. It is important to calibrate all the equipment e.g. Pipettes, Readers, Washers and/or the automated instruments used with this device, and to perform routine preventative maintenance. Interpretation
    1. Measurements and interpretation of results must be performed by a skilled individual or trained professional.

    2. Laboratory results alone are only one aspect for determining patient care and should not be the sole basis for therapy, particularly if the results conflict with other determinants.

    3. For valid test results, adequate controls and other parameters must be within the listed ranges and assay requirements.
    4. If the kits are altered, such as by mixing parts of different kits, which could produce false test results, or if results are incorrectly interpreted, shall have no liability.
    5. If computer controlled data reduction is used to interpret the results of the test, it is imperative that the predicted values for the calibrators fall within 10 % of the assigned concentrations.
    6. Total serum cortisol values may be dependent upon conditions such as time of the day for sampling or administration of prednisolone or prednisone (structurally related to cortisol). Caution must be exercised while interpreting cortisol levels for patients undergoing therapy with these and other structurally related corticosteroids such as cortisone or corticosterone. EXPECTED RANGES OF VALUES A study of normal adult population was undertaken to determine expected values for the Cortisol EIA Test System. The mean (R) values, standard deviations (sigma) and expected ranges (±2 sigma) are presented in Table
    1. TABLE 1 Population Morning Afternoon Adult 5-23 μg/dL 3-13 μg/dL Child 3-21 μg/dL 3-10 μg/dL Newborn 1-24 μg/dL Please note: Normal results may vary from lab to lab. It is important to keep in mind that establishment of a range of values which can be expected to be found by a given method for a population of "normal" persons is dependent upon a multiplicity of factors: The specificity of the method, the population tested and the precision of the method in the hands of the analyst. For these reasons each laboratory should depend upon the range of expected values established by the manufacturer only until an in-house range can be determined by the analysts using the method which a population indigenous to the area in which the laboratory is located.

    Note:

    1. Do not use reagents beyond the kit expiration date.

    2. Opened reagents are stable for sixty (60) days when stored at 2-8 °C. Kit and component stability are identified on the label.

    3. Above reagents are for a single 96-well microplate.

    Testpräzision
    The within and between assay precision of the Cortisol Microplate EIA Test System were determined by analyses on three different levels of pooled patient sera. The number (n), mean values (x), standard deviation (sigma) and coefficient of variation (C.V.) for each of these control sera are presented in Table 2 and Table3.
    TABLE 2:
    Within Assay Precision (Values in μL) Sample N X sigma CV Low 16 3.4 0.28 8.2 % Normal 16 14.2 0.91 6.4 % High 16 36.5 2.23 6.1 % Ci Substance Cross Reactivity Cortisol 1.0000 Androstenedione 0.0004 Cortisone 0.2300 Corticosterone 0.1800 11-Deoxycortisol 0.0550 Dexamethasone 0.0001 Progesterone 0.0002 17 alpha-OH Progesterone ND DHEA ND Estradiol ND Estrone ND Danazol ND Testosterone ND
    TABLE 3:
    Between Assay Precision (Values in μL!) Sample N X sigma CV Low 10 3.1 0.30 9.7 % Normal 10 15.1 1.06 7.0 % High 10 37.4 2.71 7.3 % * As measured in ten experiments in duplicate over a ten day period.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Target Alle Cortisol ELISA Kits anzeigen
    Cortisol
    Abstract
    Cortisol Produkte
    Substanzklasse
    Hormone
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