Proteins within a cell are often localized to specific cellular compartments, such as the nucleus, mitochondria, plasma membrane, or vesicles and their specific localization can provide crucial information about the function of the protein. An important technique in identifying novel proteins and understanding their function is subcellular fractionation. This process allows cells to be fractionated into compartment enriched fractions, often utilizing differential centrifugation. In addition to separating compartment specific fractions, subcellular fractionation drastically reduces the complexity of protein samples allowing for easier identification.
The Subcellular Fractionation kit exposes students to how research laboratories handle delicate samples for protein analysis and it serves as an advanced training in protein electrophoresis and protein analysis. This study involves the subcellular fractionation of a tissue sample into fractions enriched with nuclear, mitochondria or cytoplasm proteins. Students also learn about centrifugation techniques and differential centrifugation of samples for fractionation of cellular subcompartments. Students then use electrophoresis to analyze the fractionated cellular compartments and study how protein distribution or protein fingerprints differ between the subcellular compartments. Finally, students will develop an understanding of cell organelles and cellular organization.
This lab activity teaches the following:
- How to handle delicate biological samples.
- Use of differential centrifugation technique.
- Study subcellular fractionation of tissue samples.
- Use of protein electrophoresis.
- Study protein diversity at cellular level.
Please refer to the Product Protocol for additional equipment needed.