Anti-FITC MultiSort Kit ( #130-058-701 )

Anti-FITC MultiSort Kit

Overview

The Anti-FITC MultiSort Kit is a magnetic labeling system which allows the sorting of cells according to multiple surface markers. Cells are first stained with a fluorescein isothiocyanate (FITC)-conjugated primary antibody and subsequently magnetically labeled with Anti-FITC MultiSort MicroBeads. Following positive selection, the magnetic particles are removed from the cells by using the MultiSort Release Reagent. The cell population can then be magnetically labeled and sorted using a second marker of interest. This is achieved by labeling the cells directly or indirectly with MACS® MicroBeads.

Details

Detailed separation procedure
Cells are first stained with a fluorescein isothiocyanate (FITC)-conjugated primary antibody and subsequently magnetically labeled with Anti-FITC MultiSort MicroBeads. Following positive selection, the magnetic particles are removed from the cells by using the MultiSort Release Reagent. The cell population can then be magnetically labeled and sorted using a second marker of interest. This is achieved by labeling the cells directly or indirectly with MACS® MicroBeads.

Applications
The Anti-FITC MultiSort Kit has been used for the isolation of murine cytotoxic T cell subsets^1,2or α_E⁺CD4⁺ and α_E⁺CD4^- T cell subsets from murine spleen and lymph nodes³. The kit has also been used for the isolation of CD4⁺CCR5⁺ cells from synovial fluid mononuclear cells of patients with rheumatoid arthritis⁴ and for the separation of CD3⁺CD56⁺ NKT cells, CD3⁺CD56^- T cells and CD3^-CD56⁺ NK cells from human liver⁵. Moreover, the kit has been used for the separation of human CD4⁺CD45RO⁺ T cells into CD25⁺ and CD25^- subsets for further sorting according to CLA or CCR7 expression.⁶ Another application is the separation of murine GR-1⁺ cells from peripheral blood prior to MACS Separation into GR-1⁺CD18⁺ and GR-1⁺CD18^- subsets.⁷

Columns

MS, LS, XS, or autoMACS® Columns

Figure 1

Isolation of CD4⁺CD45RA⁺T cells from PBMCs with CD4-FITC, Anti-FITC MultiSort Kit, and CD45RA MicroBeads.

PBMCs before separation

Positively selected CD4⁺ cells

Positively selected CD4<sup>+</sup> cells

CD4⁺CD45RA⁺cells isolated from pre-selected CD4⁺ cells

CD4<sup>+</sup>CD45RA<sup>+ </sup>cells isolated from pre-selected CD4<sup>+</sup> cells