Description
Background information
CD3 is expressed on all T cells and CD56+ NK T cells and is associated with the T cell receptor (TCR) heterodimer. Most of the human peripheral blood lymphocytes (70–80%) and thymocytes (65–85%) are CD3+. The epitope recognized by the REAlease® CD3-Biotin Complex is located on the CD3ε chain.
Detailed separation procedure
One entire buffy coat is labeled with StraightFrom® Buffy Coat REAlease® CD3-Biotin Complex followed by Anti-Biotin MicroBeads and loaded onto Whole Blood Columns, which are placed in the magnetic field of a MultiMACS™ Cell24 Separator Plus. After removing the column from the magnetic field, the magnetically retained CD3+ cells are eluted as the positively selected cell fraction bead-free, or free of MicroBeads. Optionally, the REAlease Biotin Complex can be removed for label-free target cells.
Downstream applications
The purified CD3+ cells are bead-free, practical for consecutive magnetic isolation and optionally label-free with epitopes available for any downstream application. The isolated cells are suited for further flow cytometric analyses, molecular biology applications, and functional studies.
Overview
The StraightFrom® Buffy Coat CD3 REAlease® MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD3+ cells, starting directly from one buffy coat using the MultiMACS™ Cell24 Separator Plus for semi-automated separation or a MACS® Separator for manual separation. No sample preparation is required, including density gradient centrifugation, erythrocyte lysis, wash steps, or cell count. Whole Blood Columns are included in the kit to allow direct processing of an entire buffy coat with a simple and quick protocol.
