5 Questions
______ is a hybridization technique by which a macromolecule such as DNA, RNA, or protein is resolved in a gel matrix, transferred to a solid support, and detected with a specific probe
Blotting
______ (1981) is an Immunoblotting technique which rely on the specificity of binding between a protein of interest and a probe (antibody raised against that particular protein) to allow detection of the protein (cell, tissue, organ, or body fluid) of interest in a mixture of many other similar molecules
Western blotting
Sample preparation All sources of protein, from whole tissues, biological fluids and ______ secreted in vitro, are open to analysis by Western blotting. Assorted detergents, salts, and chemical lysis buffers such as Tris-HCI to disrupt the cellular membranes and solubilize the target protei
proteins
Sample preparation
______ 3. Blotting (or transfer) 4. Blocking 5. Antibody Probing 6. Detection 1. Sample preparation
Gel Electrophoresis
Study Notes
Western Blotting Technique
- Western blotting is a hybridization technique used to resolve and detect macromolecules like DNA, RNA, or proteins in a gel matrix, transferred to a solid support.
Immunoblotting
- Developed in 1981, Immunoblotting is a Western blotting technique that relies on the specificity of binding between a protein of interest and a probe (antibody) to detect the protein in a mixture of similar molecules.
Sample Preparation for Western Blotting
- Various sources of protein can be analyzed using Western blotting, including whole tissues, biological fluids, and secreted proteins in vitro.
- Sample preparation involves disrupting cellular membranes and solubilizing target proteins using detergents, salts, and chemical lysis buffers like Tris-HCI.
Western Blotting Protocol
- The Western blotting protocol consists of six steps:
- Sample preparation
- Blotting (or transfer)
- Blocking
- Antibody probing
- Detection
Test your knowledge of blotting techniques with this quiz. Explore concepts such as DNA, RNA, and protein resolution in gel matrices, transfer to solid supports, and detection with specific probes. Delve into the specifics of Western blotting and its reliance on the specificity of binding between proteins and probes.
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