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    12a. ELISA

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    Akbar Wibriansyah
    ELISA (Enzyme Linked Immuno-Sorbent Assay) is an immunological test used to detect the presence and quantity of a target protein. It works by detecting the binding of an enzyme-labeled antibody to the target protein. This binding allows for the qualitative detection of the target protein's presence and the quantitative measurement of how much is present. The ELISA test uses microtiter plates to capture antibodies or antigens. Detection antibodies linked to enzymes are added, followed by substrates that change color when activated by the enzymes, indicating the presence of the target. There are different ELISA methods depending on whether the capture is an antibody or antigen and whether detection is direct or indirect.

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    12a. ELISA

    Uploaded by

    Akbar Wibriansyah
    41 views13 pages
    ELISA (Enzyme Linked Immuno-Sorbent Assay) is an immunological test used to detect the presence and quantity of a target protein. It works by detecting the binding of an enzyme-labeled antibody to the target protein. This binding allows for the qualitative detection of the target protein's presence and the quantitative measurement of how much is present. The ELISA test uses microtiter plates to capture antibodies or antigens. Detection antibodies linked to enzymes are added, followed by substrates that change color when activated by the enzymes, indicating the presence of the target. There are different ELISA methods depending on whether the capture is an antibody or antigen and whether detection is direct or indirect.

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    ELISA (Enzyme Linked Immuno-Sorbent Assay) is an immunological test used to detect the presence and quantity of a target protein. It works by detecting the binding of an enzyme-labeled antibody to the target protein. This binding allows for the qualitative detection of the target protein's presence and the quantitative measurement of how much is present. The ELISA test uses microtiter plates to capture antibodies or antigens. Detection antibodies linked to enzymes are added, followed by substrates that change color when activated by the enzymes, indicating the presence of the target. There are different ELISA methods depending on whether the capture is an antibody or antigen and whether detection is direct or indirect.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
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    Download as ppt, pdf, or txt
    41 views13 pages

    12a. ELISA

    Uploaded by

    Akbar Wibriansyah
    ELISA (Enzyme Linked Immuno-Sorbent Assay) is an immunological test used to detect the presence and quantity of a target protein. It works by detecting the binding of an enzyme-labeled antibody to the target protein. This binding allows for the qualitative detection of the target protein's presence and the quantitative measurement of how much is present. The ELISA test uses microtiter plates to capture antibodies or antigens. Detection antibodies linked to enzymes are added, followed by substrates that change color when activated by the enzymes, indicating the presence of the target. There are different ELISA methods depending on whether the capture is an antibody or antigen and whether detection is direct or indirect.

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    © All Rights Reserved
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    ELISA

    Enzyme Linked Immuno-Sorbent Assay

    Heri
    Wibowo

    Definisi
    ELISA Test yang dirancang berdasarkan
    prinsip imunologi (Antigen antibodi)
    mengunakan label enzim yang dapat
    ditujukan untuk Mendeteksi keberadaan
    protein target,
    Pengikatan enzim / labeling
    memungkinkan untuk
    Melacak keberadaan protein target (Kwalitatif)
    Berapa banyak jumlah protein target
    (Kwantitatif)

    Prinsip imunologi
    Antigen
    Epitop

    Antibody

    Komponen ELISA
    Plate mikrotiter /Solid phase portion of
    assay.
    Konjugate (suatu ab yang telah berlabel
    enzim)
    Subtrate (Substansi yang akan
    mengatifkan enzim setelah berikatan dg
    bagian aktif enzim perubahan warna)
    Bloking
    Bufer pencuci/deterjen (Washing buffer)

    Metode ELISA
    Dapat didesain berdasarkan pengikat
    targetnya :
    Antigen Capture
    Antibody Capture

    Prosedur deteksinya
    Direct ELISA : Bila konjugate (Berlabel enzim)
    berikatan langsung dengan protein target /
    antigen
    Indirect ELISA : Bila konjugate (Berlabel enzim)
    tidak berikatan langsung dengan protein target
    / antigen

    Protokol Umum ELISA


    Deteksi Antibodi

    Coating
    Blocking
    Conjugate
    Subtrate

    Direct Antigen Capture

    Sumur plate Elisa

    Konjugate (Antibodi berlabel enzim


    pendeteksi
    Antigen (Target)

    Direct antibody capture

    Sumur plate Elisa


    Konjugate (Antibodi berlabel enzim
    pendeteksi
    Antigen (Target)
    Antibodi penangkap
    (Capture Ab)

    Indirect Triple Antibody


    Sandwich
    Indirect antibody capture

    Konjugate (Antibodi berlabel enzim


    pendeteksi
    Sumur plate Elisa
    Antibodi sekunder
    Antigen (Target)
    Antibodi penangkap
    (Capture Ab)

    Indirect Antigen Capture

    Konjugate (Antibodi berlabel enzim


    pendeteksi
    Sumur plate Elisa
    Antibodi target
    Antigen penangkap
    (Capture antigen)

    COMPLEMENT
    FIXATION

    Complement Fixation
    Principle

    In the complement fixation reactin an antigenantibody reaction is demonstrated by binding of


    complement.

    The reaction can be visualized with the aid of an


    indicator reactin (hemolytic system).

    The hemolytic system consist of sheep erythrocytes


    and antiserum to sheep erythrocytes (amboceptor).

    If no antigen-antibody reaction develops with the


    consumption of complement, the erythrocyte will be
    lysis completely and the hemoglobin will be
    released.

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