Table 1. Cell Membrane Isolation from Nucleus-Free Cells^a.

cell type	hypotonic buffer (ice cold)	disruption technique (ice cold)	method of isolation (4 °C)	ref
RBCs	0.25× PBS	hypotonic treatment; centrifuge 3–4 times to get pink pellet	800g, 5 min	(62)
	0.25× PBS, 0.2 mM EDTAK2		10,000 rpm, 5 min	(211)
	deionized water, EDTA		4000 rpm, 10 min	(210)
			4800 rpm, 20 min
platelets	1× PBS, 1 mM EDTA, protease inhibitor cocktail	repeated freeze thaw cycles	4000g, 3 min	(93)
	–	7 freeze–thaw cycles and sonication	discontinuous sucrose gradient ultracentrifugation	(213)
			5%, 40%, 55%
			cell membrane: interface between 5% and 40%
	deionized water	3 freeze–thaw cycles	21,000g, 7 min	(212)

bacteria	ultrafiltration	method of OMVs or EVs isolation	ref
E. coli	4000g for 10 min, filtration through a 0.45 μm vacuum filter, concentration of filtrate using ultrafiltration with a 100 kDa amicon centrifugal filter	150,000g, 2 h	(94)
S. aureus	10,000g for 20 min, filtration through a 0.45 μm vacuum filter, concentration of filtrate using ultrafiltration with a 100 kDa amicon centrifugal filter	150,000g, 3 h	(192)
K. pneumoniae	3220g for 15 min, filtration through a 0.22 μm vacuum filter, concentration of filtrate using a 100 kDa ultrafiltration tube	150,000g, 1 h	(193)

^aAbbreviations: RBC, red blood cells; PBS, phosphate-buffered saline; EDTA, ethylenediaminetetraacetic acid; EDTAK2, dipotassium EDTA; E. coli, Escherichia coli; S. aureus, Staphylococcus aureus; K. pneumonia, Klebsiella pneumonia; g, relative centrifugal force; rpm, revolutions per minute; min, minutes, h, hour; OMVs, outer membrane vesicles; EVs, extracellular vesicles.