Minute™ Plasma Membrane Protein Isolation and Cell Fractionation Kit

Minute™ Plasma Membrane Protein Isolation and Cell Fractionation Kit
is a novel and patented native PM protein isolation kit. The principle of isolation is: Cells/Tissues are first sensitized by buffer A then pass through a filter that allows cells to pass through with a zigzag path. The cell membranes are ruptured during the process and leave nuclei intact therefore the final PM prep is basically free of nuclear membrane and nuclear protein contamination. PM is separated from a mixture of un-ruptured cells (very small amount), nuclei, cytosol and organelles by subsequent differential centrifugation and density centrifugation without using ultracentrifugation. Since cell membranes are ruptured during the zigzag path when high speed centrifugal force is applied there is no need to use a homogenizer that are required by other kits. When a homogenizer is used in PM isolation, inconsistency is a major problem because variation in the duration of homogenization results in a different protein profile every time therefore resulting in a significant variation in final PM purity (inter-experiment variation). As a comparison, we use the same amount of starting cell, defined centrifugal force and pre-defined duration in every experiment and the result is much more consistent. The procedure can be completed in less than 45 min.