Subcellular Fractionation Process

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What is the purpose of subcellular fractionation?

To increase enzymic degradation in the cell

To study the whole cell under a microscope

To isolate one type of subcellular organelle from the rest of the cell contents (correct)

To identify all the enzymes present in the cell

How are cells initially broken open in subcellular fractionation?

Freezing at -80°C

Homogenization in a blender or homogenizer (correct)

Boiling in water

Shaking vigorously by hand

Why is subcellular fractionation usually carried out at 4°C?

To prevent contamination of the sample

To increase the size of subcellular organelles

To speed up the process of fractionation

To minimize enzymic degradation of the cell’s constituents (correct)

What is the purpose of straining the sample through muslin in subcellular fractionation?

To remove larger lumps of material Signup and view all the answers

How are various subcellular organelles separated in differential velocity centrifugation?

On the basis of their size Signup and view all the answers

What forces are used to separate subcellular organelles in a centrifuge during differential velocity centrifugation?

Powerful centrifugal forces Signup and view all the answers

What is the purpose of fluorescence-activated cell sorter (FACS)?

To separate cells based on fluorescence Signup and view all the answers

How are primary cultures different from secondary cultures?

Primary cultures are prepared directly from tissues. Signup and view all the answers

What is the main goal of subcellular fractionation?

To break open cells and separate organelles Signup and view all the answers

In differential velocity centrifugation, what separates the subcellular organelles?

Density Signup and view all the answers

What organelles pellet at lower forces in a differential velocity centrifugation?

Nuclei, mitochondria, chloroplasts, lysosomes Signup and view all the answers

What principle is used in equilibrium density-gradient centrifugation to separate organelles?

Density difference Signup and view all the answers

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 Signup and view all the answers

Which cellular component is pelleted first during the fractionation process?

Nuclei Signup and view all the answers

Which of the following centrifugation conditions would pellet mitochondria, peroxisomes, and chloroplasts (if present)?

6000g for 8 minutes Signup and view all the answers

What is the final supernatant fraction obtained after the complete fractionation process?

Cytosolic fraction Signup and view all the answers

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 Signup and view all the answers

Which centrifugation step separates ribosomal subunits from the cytosolic fraction?

100,000g for 90 minutes Signup and view all the answers