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        Competitive ELISA

        互联网

        1255

        DAY 1

        1. Coat Nunc immuno-module plates overnight, in usual manner.

        2. Set up competition assay between antibody and competing substance :-

        Prepare a 1:2 serial dilution of the competitive substance from 480 to 0.4 ng/ml. Add 50ul of each of the above dilutions to 25ul of 3x strength titre of antibody (i.e. a solution 3 x the titre giving 50% inhibition on direct ELISA), in sealed tubes.

        3. Incubate overnight at room temperature.

        4. Set up competition between unknown concentration of competitive substance and antibody as above.

        DAY 2

        1. Wash plate with 4x 200ul/well PBS/Tween.

        2. Add 50ul/well of antibody and incubate for 2hrs.

        3. Wash 4x with 200ul/well PBS/Tween.

        4. Add 50ul/well secondary antibody and incubate for 2hrs.

        5. Wash 4x with 200ul PBS/Tween.

        6. Add 50 ul peroxidase substrate (TMB) as for direct ELISA.

        7. Read OD @ 450nm.

        8. Plot standards against absorbance and determine unknown concentrations.

        ELISA相关仪器试剂设备

        • 酶标仪
        • 洗板机
        • 化学发光检测仪
        • ELISA检测试剂盒
        • ELISPOT检测
        • ELISA试剂盒
        • 抗体
        • 酶标板


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