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    SDS Page 1 & 2D

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    maittt.22ba13211

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    SDS Page 1 & 2D

    Uploaded by

    maittt.22ba13211
    0 views21 pages

    Original Title

    SDS PAGE 1 & 2D

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    0 views21 pages

    SDS Page 1 & 2D

    Uploaded by

    maittt.22ba13211

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 21

    Sodium Dodecyl Sulfate-Polyacrylamide gel

    Electrophoresis (SDS-PAGE)
    1. Catalase, cytochrome C,
    a-lactalbumin
    2. Hemoglobin, Cytochrome C, a- 1 2 3 4 5 6
    lactalbumin
    3. BSA, cytochrome C,
    a-lactalbumin
    4. Hemoglobin, myoglobin,
    a-lactalbumin
    5. Ferritin, cytochrome C,
    a-lactalbumin
    6. Ferritin, myoglobin,
    lighter
    a-lactalbumin
    SDS-PAGE

    Protein separation based on their molecular weight

    SDS: protein having negative charge, used to


    denature proteins
    How SDS-PAGE works

    - SDS-treated-proteins have uniformed structure and charge  the separation will depend on
    their molecular weight only.

    - Small proteins migrate faster through the gel under the influence of the applied electric field,
    whereas large proteins move slower, due to the sieving effect of the gels.
    Polyacrylamide gel

    - The polyacrylamide gel is formed by co-polymerization of acrylamide and a cross-


    linking by bis-acrylamide “.
    - Smaller pore than agarose thanks to smaller size of proteins than DNA
    - Average size of protein: 35-50 kDa
    - Average size of DNA: 2000 kDa

    -To polymerize the gel a system, consisting of ammonium persilfate (APS) (initiator) and
    tetramethylene ethylene diamin (TEMED) is added[catalyst].
    Stacking gel ad separation gel
    Concentration of stacking gel is smaller than separation gel
    What if 2 proteins have the same molecular weight?
    2D gel electrophoresis
    pH
    3 10

    High mol. Wt.

    SDS PAGE

    Low mol. Wt.

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