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The prominent tuberosity is detached using a 1 cm osteotome.
Then, the tendon is detached using Wescott scissors (fig 1C).
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Macrophages were detached using macrophage detachment medium (Promocell).
Migrated cells were detached using a detachment buffer provided with the kit and their number was quantified using a cell-permeable fluorescent dye.
After incubation of MDA-MB231 cancer cells with monocytes, as described above, cells were detached using nonenzymatic detachment solution (Sigma, Saint Quentin Fallavier, France).
Primary human monocyte-derived Mφ were detached using a 30 min incubation with Accutase™ enzyme cell detachment medium (eBioscience).
Cells were detached using 0.02%.
In experiments assessing cell viability, after treatment cells were detached using trypsin-EDTA (ThermoFisher), diluted into trypan blue, and then percentage of stained cells analyzed using an automated cell counter (TC20, Bio-Rad).
To perform binding assay, LOX-1 transfected and non-transfected CHO cells were detached using trypsin (CDS, Sigma), harvested and suspended in F12-K medium supplemented with 10% FBS.
After another 24 h cells were detached using TrypLE Express (Invitrogen) and distributed onto 3 wells of a 6-well plate with already attached irradiated feeder cells (3 × 105 cells per well irradiated with 30 Gy) in hiPS cell medium.
Cells were detached using 0.25% Tryspin/1 mM EDTA solution and added to corresponding tubes containing media.
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