Lab 8 Western

Lab 8 Western - Lab #8 Western Blot of Proteins (last...

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Unformatted text preview: Lab #8 Western Blot of Proteins (last lab!!) Objectives: To understand the concept of Western Blot. To detect the protein of interest Western Blot (immunoblotting) nitrocellulose membrane Last Week This Week Y Y Y Y Y : mouse anti- myosin (primary antibody) : goat anti-mouse conjugated to HRP Y Antibody Structure consists of four chains two heavy chains (blue) two light chains (pink) Polyclonal and monoclonal antibodies Polyclonal: recognizes several epitopes on an antigen heterogeneous Monoclonal: recognizes one epitope on an antigen. Isolated from a clone of cells producing a single, identical antibody. Generating monoclonal antibodies 1. Fuse antibody-producing cell line with immortal myeloma line. 1. Select hybrid cell for a specific antigen-antibody interaction. 1. Positive cells can be grown in culture, inject to mice or frozen. Chicken myosin protein antigen Direct vs. Indirect Detection of Antigen Direct Detection : label 1 o antibody directly...
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Lab 8 Western - Lab #8 Western Blot of Proteins (last...

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