Indicated membrane fractions obtained from the membrane fractionation were incubated with cytosol for the LC3 lipidation assay.
Alternatives derived from thermal, mechanical and enzymatic processing and from fractionation techniques by means of membranes are presented which can be exploited for fractionation and compositional matrix design of dairy products.
DOI: http://dx.doi.org/10.7554/eLife.00947.015 The data from membrane fractionation indicate that an ERGIC-enriched membrane fraction induces LC3 lipidation.
With proper purification and fractionation techniques, the contents of a particular fraction will correspond to a particular organelle.
One solution is the use of subcellular fractions such as mitochondria, nuclear and plasma membrane which can further separate from brain lysates with chemical or physical fractionation techniques.
However, only about 2% of the entries on the predicted diatom proteome database have been identified in this study, suggesting that further development of extraction and fractionation techniques are required to improve the identification of low abundant or membrane proteins.
Using sub-organellar fractionation techniques, both with and without detergents, combined with electron microscopy, we found that both the limiting membrane of the organel and the intraluminal vesicles contain raft-type membrane domains.
Plasma membrane fractionation was performed using a membrane protein extraction kit from BioVision.
Outer membrane fractionation confirmed that the processed protein was located in the outer membrane.
Thermal fractionation provides an alternative to experimentally more time consuming and complicated fractionation techniques that involve preparative or analytical fractionation in solution.
