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MACS® Technology offers various separation strategies, which provide the perfect basis for the isolation of almost any cell type from any species and various sample materials.
MACS Cell Separation Strategies poster
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| Positive selection |
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Depletion |
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Untouched isolation |
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Sequential sorting |
The fastest way to pure target cells
Positive selection means that a particular target cell type is magnetically labeled. During separation, the magnetically labeled cells are retained within the column. Unlabeled cells flow through. After a washing step, the column is removed from the magnetic field of the separator, and the target cells are eluted from the column.
Positive selection can be performed by direct or indirect magnetic labeling.
Specific MACS MicroBeads are available for the positive selection of numerous cell types.
Removal of an unwanted cell type
To remove a certain cell type from a mixture of cells, the unwanted cell type is magnetically labeled. During separation, the unlabeled target cells are collected in the flow-through fraction. The unwanted cell type is retained within the column.
Optionally, the retained cells can be eluted after removal of the column from the separator (not shown).
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Isolation of cells in an unlabeled form
To isolate a particular target cell type in an unlabeled, i.e., untouched form, non-target cells are magnetically labeled and depleted. During separation, the unlabeled target cell type is collected in the flow-through fraction. The mixture of magnetically labeled non-target cells is retained within the column.
Optionally, the magnetically labeled cells can be eluted after removal of the column from the magnetic field of the separator (not shown).
MACS Cell Isolation Kits for untouched isolation contain a cocktail of titrated antibodies and MACS MicroBeads for indirect magnetic labeling. They are the preferred choice if binding of antibodies to the target cells is not desired.
Isolation of a cell subset by using more than one marker
A combination of two subsequent separations is applied to isolate cell subsets that can be distinguished from other cell types through their expression of two different markers. This includes cell types for which a specific marker has not been defined.
Depletion followed by positive selection:
This separation strategy is useful, if undesired cells and target cells have one marker in common. In this case, the target cells cannot be isolated in a single positive selection step using this marker.
Therefore, the undesired cells expressing this marker are magnetically labeled via antigens distinct from the common marker, and depleted. The cells appearing in the flow-through fraction during this separation are subsequently labeled with MACS MicroBeads that bind to the common marker. Target cells are then isolated by positive selection.
Sophisticated MACS Cell Isolation Kits based on this strategy are available for the fast and convenient isolation of specific cell subsets.
Two subsequent positive selections:
Multiparameter sorting with MACS MultiSort MicroBeads is based on two sequential positive selections according to two different markers.
MACS MultiSort MicroBeads specific for the first marker allow the first positive selection. After the separation, the cells are incubated with the MultiSort Release Reagent, which enzymatically removes the MultiSort MicroBeads from the cells. In the next step, the target cells are magnetically labeled with MACS MicroBeads directed against the second marker and again subjected to positive selection.
Isolation of a cell subset by using more than one marker
A combination of two subsequent separations is applied to isolate cell subsets that can be distinguished from other cell types through their expression of two different markers. This includes cell types for which a specific marker has not been defined.
Depletion followed by positive selection:
This separation strategy is useful, if undesired cells and target cells have one marker in common. In this case, the target cells cannot be isolated in a single positive selection step using this marker.
Therefore, the undesired cells expressing this marker are magnetically labeled via antigens distinct from the common marker, and depleted. The cells appearing in the flow-through fraction during this separation are subsequently labeled with MACS MicroBeads that bind to the common marker. Target cells are then isolated by positive selection.
Sophisticated MACS Cell Isolation Kits based on this strategy are available for the fast and convenient isolation of specific cell subsets.
Two subsequent positive selections:
Multiparameter sorting with MACS MultiSort MicroBeads is based on two sequential positive selections according to two different markers.
MACS MultiSort MicroBeads specific for the first marker allow the first positive selection. After the separation, the cells are incubated with the MultiSort Release Reagent, which enzymatically removes the MultiSort MicroBeads from the cells. In the next step, the target cells are magnetically labeled with MACS MicroBeads directed against the second marker and again subjected to positive selection.
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