EasySep Direct Monocyte-RoboSep

From STEMCELL Technologies, this immunomagnetic negative selection kit isolates highly purified CD14+ monocytes directly from human whole blood, with no requirement for density gradient centrifugation, sedimentation, or lysis. The kit provides greater than 99.9% red blood cell depletion and achieves up to 92% purity of isolated cells. Isolated cells are untouched and ready for flow cytometry, functional assays, culture, or nucleic acid extraction immediately following the procedure.

The method targets non-monocytes and CD16+ monocytes for removal using a cocktail of monoclonal antibodies together with magnetic particles (RapidSpheres™); desired cells are collected after separation in a compatible EasySep™ magnet, including the EasySep™ Magnet, "The Big Easy", Easy 50, EasyEights™, or the automated RoboSep™. The kit is designed for processing 100 mL of whole blood. EDTA (K2EDTA or K3EDTA) is required for performance, and unprocessed whole blood less than 24 hours old is recommended for best recovery.

Typical monocyte (CD14+) content of the non-lysed final isolated fraction from healthy donor whole blood is 82.2 ± 8.4% (gated on CD45) or 79.0 ± 10.1% (not gated on CD45). The starting frequency of monocytes in the non-lysed whole blood sample is approximately 0.007% (data not shown). Kit components include an isolation cocktail (a combination of monoclonal antibodies in PBS with an Fc receptor blocking antibody) and RapidSpheres™ (a suspension of magnetic particles and monoclonal antibodies in PBS); both are stored at 2 - 8°C and must not be frozen.