Diamond CD133 Isolation Kit ( #130-094-913 )

Diamond CD133 Isolation Kit

Overview

The Diamond CD133 Isolation Kit was specifically developed for the isolation of highly pure Lin^–CD133⁺ stem cells from peripheral blood, cord blood, bone marrow, and leukapheresis product by sequential sorting.

Details

Detailed separation procedure
The Diamond CD133 MicroBead Kit includes a lineage depletion followed by a CD133 isolation. In the first step, cells are labeled with a cocktail of biotin-conjugated antibodies against lineage-specific antigens. These cells are subsequently labeled with Anti-Biotin MicroBeads. Lineage-negative stem and progenitor cells are obtained by depletion of the magnetically labeled cells. Then CD133⁺ cells are isolated from the lineage-negative stem and progenitor cells using CD133 MicroBeads. The resulting population obtained are Lin^-CD133⁺stem cells.

Diamond CD133 MicroBeads recognize the epitope CD133/1. For control staining of separated cells, the application of CD133/2 (293C3) antibody is recommended.

Downstream applications
Lin^-CD133⁺stem cells can be used for many applications, including:

gene or miRNA expression profiling
characterization of undifferentiated stem cells and their developmental potential
stimulation of proliferation and maturation by cytokines
HSC-CFU assays
adoptive transfer and hematopoietic reconstitution studies

Columns

For the first magnetic separation (depletion): LD or autoMACS Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns

Figure 1

CD133⁺Lin^– cells were isolated from human PBMCs using the Diamond CD133 Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator, and a MiniMACS™ Separator. Cells were fluorescently stained with CD34-FITC, CD133/2 (293C3)-PE, CD45-APC, and Anti-Biotin-PE and analyzed using the MACSQuant® Analyzer.

PBMCs before separation

Lineage-negative fraction

$Lineage-negative fraction$

Isolated CD133⁺Lin^- cells

Isolated CD133<sup>+</sup>Lin<sup>-</sup> cells