18 Questions
What is the purpose of subcellular fractionation?
To isolate one type of subcellular organelle from the rest of the cell contents
How are cells initially broken open in subcellular fractionation?
Homogenization in a blender or homogenizer
Why is subcellular fractionation usually carried out at 4°C?
To minimize enzymic degradation of the cell’s constituents
What is the purpose of straining the sample through muslin in subcellular fractionation?
To remove larger lumps of material
How are various subcellular organelles separated in differential velocity centrifugation?
On the basis of their size
What forces are used to separate subcellular organelles in a centrifuge during differential velocity centrifugation?
Powerful centrifugal forces
What is the purpose of fluorescence-activated cell sorter (FACS)?
To separate cells based on fluorescence
How are primary cultures different from secondary cultures?
Primary cultures are prepared directly from tissues.
What is the main goal of subcellular fractionation?
To break open cells and separate organelles
In differential velocity centrifugation, what separates the subcellular organelles?
Density
What organelles pellet at lower forces in a differential velocity centrifugation?
Nuclei, mitochondria, chloroplasts, lysosomes
What principle is used in equilibrium density-gradient centrifugation to separate organelles?
Density difference
What is the purpose of using increasingly higher g forces for longer periods during cell fractionation?
To separate larger cellular components first, followed by smaller ones
Which cellular component is pelleted first during the fractionation process?
Nuclei
Which of the following centrifugation conditions would pellet mitochondria, peroxisomes, and chloroplasts (if present)?
6000g for 8 minutes
What is the final supernatant fraction obtained after the complete fractionation process?
Cytosolic fraction
Which of the following cellular components would be found in the pellet obtained after centrifugation at 40,000g for 30 minutes?
Plasma membrane and Golgi fragments
Which centrifugation step separates ribosomal subunits from the cytosolic fraction?
100,000g for 90 minutes
Learn about the process of isolating subcellular organelles from cells through subcellular fractionation, which involves rupturing the plasma membrane by homogenization in an isotonic sucrose solution. This method helps in studying macromolecules and metabolic processes within cells.
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