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  • Western Blot

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    Western blotting is a technique used to detect specific proteins in a sample of tissue or cells. It involves lysing cells to release proteins, determining the protein concentration, loading equal amounts of protein sample onto a gel, and running the gel to separate proteins by molecular weight. A loading control protein is also run to ensure equal loading of protein samples in each lane.

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    Western blotting is a technique used to detect specific proteins in a sample of tissue or cells. It involves lysing cells to release proteins, determining the protein concentration, loading equal amounts of protein sample onto a gel, and running the gel to separate proteins by molecular weight. A loading control protein is also run to ensure equal loading of protein samples in each lane.

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    3 views8 pages

    Western Blot

    Uploaded by

    api-438165055
    Western blotting is a technique used to detect specific proteins in a sample of tissue or cells. It involves lysing cells to release proteins, determining the protein concentration, loading equal amounts of protein sample onto a gel, and running the gel to separate proteins by molecular weight. A loading control protein is also run to ensure equal loading of protein samples in each lane.

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    © All Rights Reserved
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    Western Blotting

    Sample Prep
    • Collection of tissue
    • Lysis Buffer
    • Breaks open cells
    • Protease Inhibitors
    • Sonication
    • Centrifugation
    Protein Concentration
    • We need to know how many mg/mL of protein we have in our samples.
    With this information we will know how many uL of sample to use in the
    western blot.
    • Why?
    • Create standards
    • Collect absorbance readings
    • Create standard curve and determine line
    of best fit.
    • Determine concentration of your samples.
    Practice Worksheet
    Running the western
    • Once you have the protein concentrations calculated, you can load
    the gel with equal amount of protein in each samples.
    • http://www.abcam.com/protocols/general-western-blot-protocol
    Loading Control

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