YILIMART - StraightFrom BuffyCoat CD3 MB Kit

The StraightFrom

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Buffy Coat CD3 MicroBead Kit has been developed for the rapid positive selection of CD3

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cells directly from one buffy coat using the MultiMACS™ Cell24 Separator Plus for semi-automated separation or a MACS

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Separator for manual separation. No sample preparation is required, including density gradient centrifugation, erythrocyte lysis, wash steps, or cell count. Whole Blood Columns and Whole Blood Column Elution Buffer are included in the kit to allow direct processing of an entire buffy coat.

Background information

The CD3 antigen is associated with the T cell receptor (TCR) heterodimer. CD3 is expressed on all T cells, constituting 15–20% of human peripheral blood leukocytes.

Detailed separation procedure

One entire buffy coat is labeled with StraightFrom

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Buffy Coat CD3 MicroBeads and loaded onto Whole Blood Columns, which are placed in the magnetic field of a MultiMACS™ Cell24 Separator Plus or a MACS

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Separator. After removing the column from the magnetic field, the magnetically retained CD3

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cells can be eluted as the positively selected cell fraction.

Downstream applications

The purified CD3

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cells are well suited for further flow cytometric, functional, or molecular analysis.

Figure 1

Separation of a buffy coat sample using the StraightFrom

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Buffy Coat CD3 MicroBead Kit and the MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter and Whole Blood Columns. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD14-VioGreen™, as well as with CD45-VioBlue

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and analyzed by flow cytometry using the MACSQuant

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Analyzer. Viable leukocytes were gated for analysis based on scatter signals, propidium iodide fluorescence, and CD45 expression.