YILIMART - Anti-Sca-1 MicroBeads, mouse

The Anti-Sca-1 MicroBeads (non-HSC) have been developed for the positive selection of non-hematopoietic stem and progenitor cells from various tissues. For the isolation of HSCs based on Sca-1 expression the Anti-Sca-1 MicroBead Kit (VioBright™ FITC), mouse should be used.

Background information

Sca-1 (stem cell antigen-1) is a 18 kDa GPI-linked surface protein of the Ly-6 family (Ly-6A/E). The Anti-Sca-1 antibody clone D7 specifically recognizes both Ly-6E.1 and Ly-6A.2, which are gene products of the two Ly-6A/E alleles expressed in different mouse strains (Ly-6E.1: e.g. BALB/c, C3H, NZB; Ly6A.2: e.g. C57BL/6, SJL, 129, AKR).

Sca-1 is expressed on hematopoietic stem cells and progenitor cells in mouse bone marrow and one of the defining markers for so-called KSL (c-kit

⁺

Sca-1

⁺

Lin

^-

) cells. Sca-1 in combination with CD105 (Endoglin) expression has been used to define long-term repopulating hematopoietic stem cells (LTR-HSCs) in mouse bone marrow. In addition, Sca-1

⁺

bone marrow cells also contain the mesenchymal stem cell fraction present. Sca-1

⁺

cells have been shown to give rise to hepatocytes

in vivo

and neural cells

in vitro

. Furthermore, Sca-1 is expressed on stem cells in a variety of nonhematopoietic tissues, such as adult liver, heart, and prostate. Anti-Sca-1 MicroBeads have been developed for the separation of mouse cells based on the expression of Sca-1 except for HSCs, for which the Anti-Sca-1 MicroBead Kit (VioBright™ FITC), mouse should be used.

Figure 1

Mouse spleen was dissociated using the Spleen Dissociation Kit, mouse (# 130-095-926) and the gentleMACS™ Dissociator. Sca-1–expressing cells were isolated using Anti-Sca-1 (non-HSC) MicroBeads, an LS Column, and a QuadroMACS™ Separator. Cells were fluorescently stained with CD45-APC (# 130-110-660) and Anti-Sca-1-VioBright™ FITC (# 130-106-259) and analyzed by flow cytometry using the MACSQuant

^®

Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Before separation	Sca-1 ⁺ cells

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Anti-Sca-1 MicroBeads, mouse

For research use only

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Background information

Figure 1

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