Cell lysates were passed 10 times through a 21-gauge needle, and 400 µl of lysate were mixed with 800 µl of a 60% OptiPrep solution (Axis-Shield, Oslo, Norway) (40% final concentration).
Lymphocytes were isolated using Lymphoprep solution (Axis-Shield, Oslo, Norway).
These solutions were prepared from Optiprep iodixanol solution (Axis-Sheild, Oslo, Norway) with 60 mM Tris HCl (pH 7.5).
The microvesicles were harvested and mixed with an equal volume of 60% iodixanol solution (Axis-Shield PoC AS, Oslo, Norway), to give 30% iodixanol.
The blood was diluted 1 1 with Hank's balanced salt solution, layered on Lymphoprep™ solution (Axis-Shield, Dundee, Scotland) and centrifuged for 20 minutes at 800 g.
Human peripheral blood mononuclear cells (PBMCs) were isolated from 50 ml of whole blood obtained from a healthy female volunteer using Lymphoprep solution (Axis-shield) according to manufacturer's protocol.
The cytosol and membrane fractions were then centrifuged at 100,000 × g for 60 min. For OptiPrep gradient (Li, 2008 #19), 200 µl of lysate were mixed with 200 µl of a 50% OptiPrep solution (Axis-Shield) (25% final concentration).
In this paper, we shall establish the unilateral global bifurcation which bifurcates from the trivial solutions axis or from infinity of a class of nonlinear Sturm Liouville problems with nondifferentiable nonlinearity, respectively.
The supernatant was used for the isolation of HSCs using OptiPrep™ self-forming density gradient solutions (Axis-Shield PoC AS, Rodeløkka, Norway) as described previously [20].
The nuclear pellet was resuspended in a 30% iodixanol solution (Optiprep, Axis Shield), and centrifuged at 12,000× g for 4 min. This last centrifugation step was repeated a second time, and the pellet obtained was used as a pure nuclear fraction.
