Microsome Isolation
Microsome Isolation - Related Videos
05:09
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2023
This video describes the procedure for isolating mitochondria from human ovarian cancer tissues using differential-speed centrifugation in combination with density gradient centrifugation. The isolated mitochondria can be used for proteomic analysis of human ovarian cancer mitochondrial proteome.
09:16
Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation
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Cited by 9 •
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The goal of this protocol is to provide a cell-based system that replicates the formation of alpha-synuclein aggregates in vivo. Intracellular alpha-synuclein inclusions are seeded in primary neurons by the internalization and propagation of exogenous administered native microsomes-associated alpha-synuclein aggregates isolated from diseased alpha-synuclein transgenic mice.
10:27
Isolation and Culture of Hippocampal Neurons from Prenatal Mice
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Cited by 181 •
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We provide a protocol for the culture of highly purified hippocampal neurons from prenatal mouse brains without the use of a feeder glial cell layer.
08:09
Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice
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Cited by 1 •
2017
We describe the isolation of cardiac extracellular matrix from C57Bl/6J control mice, tight-skin mice, and tight-skin mice treated with the IRF5 inhibitory peptide. We also describe the vasodilation studies on the isolated vessels from C57Bl/6J, tight-skin mice and tight-skin mice treated with the IRF5 inhibitory peptide.
07:12
Neutrophil Isolation Protocol
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Cited by 173 •
2008
Neutrophils are among the first cells to arrive on the site of inflammatory immune response, and their functions and mechanisms have been studied extensively in vitro. We demonstrate a standard density gradient separation method to isolate human neutrophils from whole blood using commercially available separation media.
FAQs
Homogenization converts the endoplasmic reticulum into sealed vesicles rather than leaving it as intact cellular sheets. This physical change makes the membrane material compatible with fractionation while preserving membrane-associated enzymes, transport proteins, and ribosomes in a recoverable form. Consequently, investigators can examine endoplasmic-reticulum-related activities in a preparation separated from whole-cell complexity.
The fraction’s value comes from the molecular components it carries. Membrane-associated enzymes can be assayed, transport proteins can be examined as part of the isolated membrane material, and ribosomes connect the preparation to protein synthesis studies. Because these components are collected together in a controlled fraction, microsomes support analysis focused on endoplasmic-reticulum-associated functions.
Differential centrifugation separates material according to the cellular components recovered during processing. It removes larger structures such as nuclei and mitochondria before the microsomal fraction is analyzed, while ultracentrifugation can provide an additional separation step. This staged approach produces a preparation more suitable for examining membrane-associated activities than an unfractionated cell homogenate.
Isolated microsomes are especially useful when researchers need a controlled system for examining drug biotransformation. Their membrane-associated enzymes allow investigators to measure how compounds are processed and to characterize related metabolic pathways. This application helps connect enzyme activity observed in the fraction with the broader question of how substances undergo biochemical modification in cells.
A basic workflow begins with disrupted cells or tissue, followed by homogenization to break the endoplasmic reticulum into sealed vesicles. Differential centrifugation then separates the vesicle-containing material from nuclei, mitochondria, and other components. When greater separation is needed, ultracentrifugation follows, producing a microsomal preparation for biochemical measurements.
Microsomal preparations can reveal enzyme activity associated with the processing of compounds and support characterization of the metabolic pathways involved. Because the material is studied outside the complete cell, investigators obtain a controlled biochemical setting for evaluating compound processing. The resulting measurements are useful for comparing how different substances are handled by membrane-associated enzymatic systems.
In biology, the preparation links endoplasmic reticulum structure with measurable functions. Researchers can investigate membrane-associated enzymes and transport proteins while also examining processes connected with protein synthesis and lipid metabolism. Studying these activities in an isolated fraction provides a focused way to analyze endoplasmic-reticulum-related functions without relying only on observations from intact cells.