YILIMART - Anti-FITC-Multisort Kit

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.

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

^1,2

or α

_E ⁺

CD4

⁺

and α

_E ⁺

CD4

^-

T cell subsets from murine spleen and lymph nodes

³

. The kit has also been used for the isolation of CD4

⁺

CCR5

⁺

cells from synovial fluid mononuclear cells of patients with rheumatoid arthritis

⁴

and for the separation of CD3

⁺

CD56

⁺

NKT cells, CD3

⁺

CD56

^-

T cells and CD3

^-

CD56

⁺

NK cells from human liver

⁵

. 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

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

Selected references
1. Flynn, S. et al. (1998) CD4 T cell cytokine differentiation: the B cell activation molecule, OX40 ligand, instructs CD4 T cells to express interleukin 4 and upregulates expression of the chemokine receptor, Blr-1. J. Exp. Med. 188: 297-304
2. Holtappels, R. et al. (2000) Enrichment of immediate-early 1 (m123/pp89) peptide-specific CD8 T cells in a pulmonary CD62L(lo) memory-effector cell pool during latent murine cytomegalovirus infection of the lungs. J. Virol. 24: 11495-11503
3. Lehmann, J. et al. (2002) Expression of the integrin alpha Ebeta 7 identifies unique subsets of CD25+ as well as CD25- regulatory T cells. Proc. Natl. Acad. Sci. U.S.A. 99: 13031-13036
4. Suzuki, N. et al. (1999)
  
  Selective accumulation of CCR5
  ⁺
  T lymphocytes into inflamed joints of rheumatoid arthritis.
  
  Int. Immunol. 11: 553-559
5. Doherty, D. G. et al. (1999)
  
  The human liver contains multiple populations of NK cells, T cells, and CD3
  ⁺
  CD56
  ⁺
  natural T cells with distinct cytotoxic activities and Tʜ1, Tʜ2, and Tʜ0 cytokine secretion patterns.
  
  J. Immunol. 163: 2314-2321
6. Iellem et al. (2003)
  
  Skin-versus gut-skewed homing receptor expression and intrinsic CCR4 expression on human peripheral blood CD4
  ⁺
  CD25
  ⁺
  suppressor T cells.
  
  Eur. J. Immunol. 33: 1488-1496
7. Weinmann, P. et al. (2003) A role for apoptosis in the control of neutrophil homeostasis in the circulation: insights from CD18-deficient mice. Blood 101: 739-746

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Anti-FITC-Multisort Kit

For research use only

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Detailed separation procedure

Applications

Columns

Figure 1

Selected references

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