YILIMART - StraightFrom BuffyCoat CD4/CD8 MB Kit

The StraightFrom

^®

Buffy Coat CD4/CD8 MicroBead Kit has been developed for the rapid simultaneous positive selection of CD4

⁺

and CD8

⁺

cells 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 and Whole Blood Column Elution Buffer are included in the kit to allow direct processing of an entire buffy coat.

Background information

The CD4 antigen is expressed on T helper (Tʜ) cells and at a lower level on monocytes and dendritic cells. The CD4 antigen is an accessory molecule in the recognition of MHC class II/peptide complexes by the TCR heterodimer on CD4

⁺

Tʜ cells. The CD8 antigen is expressed on cytotoxic T cells and on a subset of CD16

⁺

NK cells. The CD8 antigen acts as an accessory molecule in the recognition of MHC class I/peptide complexes by the TCR heterodimer on CD8

⁺

cytotoxic T cells.

Detailed separation procedure

One entire buffy coat is labeled with StraightFrom

^®

Buffy Coat CD4/CD8 MicroBeads and loaded onto Whole Blood Columns, which are placed in the magnetic field of a MultiMACS™ Cell24 Separator Plus or a MACS

^®

Separator. After removing the column from the magnetic field, the magnetically retained CD4

⁺

and CD8

⁺

cells can be eluted as the positively selected cell fraction.

Downstream applications

The purified CD4

⁺

and CD8

⁺

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

Figure 1

Separation of a buffy coat sample using the StraightFrom

^®

Buffy Coat CD4/CD8 MicroBead Kit and the MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter and Whole Blood Columns. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE-Vio

^®

770, CD8-APC, as well as CD45‑VioBlue

^®

analyzed by flow cytometry using the MACSQuant

^®

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