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Receptor-specific T cell staining with reversible Fab Streptamers®

Fluorescent Fab Streptamers®, adducts of Strep-tagged Fab fragments (Fab-Streps) and Strep-Tactin® PE or APC, are directed against cell surface markers including those of T cells (CD3, CD4, CD8), memory T cells (CD8, CD62L, CD45RA-), regulatory T cells (CD4, CD25, CD45RA).

Cell staining procedure

1) Fab-Streps (Strep-tagged Fab fragments) are multimerized with Strep-Tactin® attached to a fluorochrome to generate multimers with high binding avidity. These multimers are directed to the respective cell surface marker(s) and are thus determined for staining of the target cell (Fig.1).

Fab Streptamer fluorochrome
Fig.1: Fab Streptamer® Fluorochrome

2) After FACS analysis or FACS sorting of the target cells, the staining reagents can be completely removed with biotin (Fig.2). Cells are left phenotypically and functionally indistinguishable from untreated cells.

Fig. 2: Target cells are selected by a Fab Streptamer®. After staining and fluorescence assisted sorting, the Strep-Tactin® complex is removed from the target cells by the addition of biotin under physiological conditions at low temperatures (4°C). The monomeric low affinity Fab-Strep molecules are then dissociated completely from the cell surface by two short washing steps. The isolated cells are phenotypically and functionally undistinguishable from non-treated cells and predestined for clinical use.

For further information about the receptor-specific T cell staining with Fab Streptamers® and the suitable reagents please have a look at our workflow.

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