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  • 产品名称：Penicillin ELISA Kit

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  • 产品厂商：Ddiagautoea
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简单介绍：
Penicillin ELISA Kit
详情介绍：
Purpose Enzyme Immunoassay for the Quantitative Determination of Penicillin in Food
Analytical Method Quantitative
Detection Method Colorimetric
Sensitivity 3 ng/mL
Research Area Drugs
Assay Time 2 - 3 h
Plate Pre-coated
Assay Procedure
  1. Prepare samples as described above.
    2. Pipet 100 μL ready-to use standards or prepared samples in duplicate into the appropriate wells of the microtiter plate. Immediately add 50 μL penicillin antibody into each well.
    3. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
    4. Wash the plate three times as follows: Discard the contents of the wells (dump or aspirate). Pipet 300 μL of diluted washing solution into each well. After the third repetition empty the wells again and remove residual liquid by striking the plate against a paper towel. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbencies.
    5. Pipet 100 μL of conjugate (anti-mouse-IgG-HRP) into each well.
    6. Cover the microtiter plate with a plastic foil and incubate for 60 minutes at room temperature on a microtiter plate shaker (or 90 minutes without shaker).
    7. Wash the plate as outlined in 4.
    8. Pipet 100 μL of substrate solution into each well.
    9. Allow the reaction to develop in the dark (e.g. cupboard or drawer, the chromogen is light-sensitive) for 20 minutes at room temperature.
    10. Stop enzyme reaction by adding 100 μL of stop solution (0.5 M H2SO4) into each well. The blue colour will turn yellow upon addition.
    11. After thorough mixing, measure absorbance at 450 nm (reference wavelength 620 nm), using an ELISA reader. The colour is stable for 30 minutes.
Calculation of Results
  1. Calculate the average optical density (OD 450 nm) for each set of reference standards or samples.

    2. Construct a standard curve by plotting the mean optical density obtained for each reference standard against its concentration in ng/mL on semi-log graph paper with the optical density on the vertical (y) axis and the concentration on the horizontal (x) axis.

    3. Using the mean optical density value for each sample, determine the corresponding concentration ' of penicillin in ng/mL from the standard curve. Depending on experience and/or the availability of ' computer capability, other methods of data reduction may be employed.
    4. The diluted samples must be further converted by the appropriate dilution factor. The dilution factor is 20 for shrimps und 4 for milk extraction according to the sample preparation procedure as described above.
    TYPICAL STANDARD VALUES
    The following table contains an example for a typical standard curve. The binding is calculated as percent of the absorption of the 0 ng/mL standard. These values are only an example and should not be used instead of the standard curve which has to be measured in every new test. Penicillin (ng/mL) ( % binding of 0 ng/mL) 0 100 4 85 10 70 40 35 100 15 400 5
Restrictions For Research Use only
Storage 4 °C
Storage Comment Store at 2-8 °C
Background publications Klosz: "Quality management for the processing of medical devices." in: GMS Krankenhaushygiene interdisziplinär, Vol. 3, Issue 3, pp. Doc22, 2010 (PubMed).

Knecht, Strasser, Dietrich, Märtlbauer, Niessner, Weller: "Automated microarray system for the simultaneous detection of antibiotics in milk." in: Analytical chemistry, Vol. 76, Issue 3, pp. 646-54, 2004 (PubMed).

Gustavsson, Sternesjö: "Biosensor analysis of beta-lactams in milk: comparison with microbiological, immunological, and receptor-based screening methods." in: Journal of AOAC International, Vol. 87, Issue 3, pp. 614-20, 2004 (PubMed).

Huth, Warholic, Devou, Chaney, Clark: "Parallux beta-lactam: a capillary-based fluorescent immunoassay for the determination of penicillin-G, ampicillin, amoxicillin, cloxacillin, cephapirin, and ceftiofur in bovine milk." in: Journal of AOAC International, Vol. 85, Issue 2, pp. 355-64, 2002 (PubMed).

Althaus, Molina, Rodriguez, Fernandez: "Detection limits of beta-lactam antibiotics in ewe milk by penzym enzymatic test." in: Journal of food protection, Vol. 64, Issue 11, pp. 1844-7, 2001 (PubMed).

de Leuw, Kapa, Petz, Schreurs, Kan: "Induction and characterization of multianalyte antibodies against penicillins in egg yolk." in: Journal of AOAC International, Vol. 80, Issue 6, pp. 1220-8, 1998 (PubMed).

Haioun, Lepage, Gisselbrecht, Coiffier, Bosly, Tilly, Morel, Nouvel, Herbrecht, DAgay et al.: "Comparison of autologous bone marrow transplantation with sequential chemotherapy for intermediate-grade and high-grade non-Hodgkin's lymphoma in first complete remission: a study of 464 patients. ..." in: Journal of clinical oncology : official journal of the American Society of Clinical Oncology, Vol. 12, Issue 12, pp. 2543-51, 1995 (PubMed).

Usleber, Lorber, Straka, Terplan, Märtlbauer: "Enzyme immunoassay for the detection of isoxazolyl penicillin antibiotics in milk." in: The Analyst, Vol. 119, Issue 12, pp. 2765-8, 1995 (PubMed).