As VE-cad is particularly sensitive to trypsin in absence of Ca2+, mono-cultured HUVEC and co-cultured HUVEC were detached in a way described before with minor modifications to maximally preserve VE-cad at the cell surface and to avoid its degradation [5].
These were detached in a 30% KOH solution, washed several times in DI water, and transferred onto TEM grids.
After being incubated in the dark for 20 min, cells were detached in a trysin-EDTA solution, spun down and washed twice in PBS.
Comparing our electrophysiology data with the confocal images, it is likely that a large number of scolopidia need to be detached in order for an electrophysiological difference to be detected with our extracellular recording.
A series of interconnected cracks appear in these situations and make the asphalt highly unstable, and pieces of asphalt can be detached in the worst cases.
Earl Spencer however demurred, arguing that as Nelson had only been detached in command of a squadron, rather than being the commander in chief of the fleet, such an award would create an unwelcome precedent.
While the main fleets manoeuvered around one another, Castor was detached in pursuit of a Dutch merchant ship and on 29 May encountered the smaller independently cruising British frigate HMS Carysfort.
After attachment, cells were treated for 24 h inhibitors (or vehicle in the controls); then the culture-insert was detached in order to form a cell-free gap into the cell monolayer.
He emerges as a bundle of contradictions: a man of immense people skills and limited policy interest, a candidate who excels at the sport of politics but is detached in the business of governing, a deep skeptic about the role of the government that he now seeks to lead.
Following six hours of treatment, cells infected with a membrane-targeted GFP were detached in drug-containing media and McTNs were scored blindly.
