20 Questions
What is the main purpose of using detergent in western blotting?
To prevent nonspecific binding of the antibody
Which type of detergent is commonly used for solubilizing most proteins by disrupting non-covalent bonds within and between proteins?
Sodium dodecyl sulfate (SDS)
What is the minimum concentration of protein extract recommended for sample preparation to avoid loss of protein and large volumes of samples to be loaded onto gels?
0.1 mg/mL
Which type of detergent is considered mild surfactants, typically not disrupting protein-protein interactions, and generally not denaturing proteins?
Non-ionic detergents
What is the purpose of adding an equal volume of 2X Laemmli sample buffer to a specific volume of protein?
To reduce and denature the proteins
What is the purpose of Western Blotting (WB) technique?
To identify specific proteins within a complex mixture
Which method is a prerequisite for Western blotting?
SDS PAGE technique
What is the composition of NP-40 buffer used in sample preparation for Western blotting?
150 mM NaCl, 1.0% NP-40, 50 mM Tris-HCl, pH 8.0, Protease inhibitors
What is the role of detergents in biomedical laboratories?
To disrupt cell membranes and release intracellular materials
What principle does Western Blotting rely on for the identification of specific proteins?
Specific binding between a protein of interest and antibody raised against that protein
Anionic detergents like sodium dodecyl sulfate (SDS) disrupt non-covalent bonds within and between proteins, denaturing them, and resulting in the loss of their native conformation and function.
True
Sodium deoxycholate and sodium cholate are non-ionic detergents.
False
Non-ionic detergents generally denature proteins and disrupt protein-protein interactions.
False
The purpose of using detergent in western blotting is primarily to solubilize proteins.
False
The amphiphilic property of detergents allows them to prevent nonspecific binding in immunochemical assays and protein crystallization.
True
The SDS PAGE technique is not a prerequisite for Western blotting.
False
NP-40 buffer contains 0.5% sodium deoxycholate.
False
RIPA buffer contains 0.1% SDS (sodium dodecyl sulphate).
True
Detergents used in biomedical laboratories are not amphiphilic molecules.
False
Western Blot (Immunoblotting) technique aims to identify DNA within a complex mixture.
False
Test your knowledge of Western Blotting (Immunoblotting) technique, which aims to identify specific proteins within a complex mixture by utilizing antibody-based methods. This quiz covers the procedures overview including sample preparation, SDS PAGE technique, and protein identification.
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