Forces required to disrupt cell wall may rapture cell organelles. The challenge of cell fractionation To chose methods of cell disruption and incubation of subcellular fractions that are appropriate to the particular investigation and which minimizes artefacts. Can be transformed into quinones. Quinones can react with reactive groups of proteins to form a dark pigment, melanin.
Melanin can inactivate many enzymes.
Can form ionic interactions with basic amino acids of proteins. Can form hydrophobic bonds with hydrophobic regions of proteins. Temperature requirements for cell fractionation Performed at 4 C to reduce the activity of proteases. All media and apparatus should be precooled and maintained at 4 C. Unwanted organelle damage may be caused by shearing forces required to break the cell wall. Enzyme activity may be lost if there is foaming of the homogenate in the high-speed blender.