MACS® Cell Separation (Manual)

MACS® MicroBead Technology is the leading method for column-based magnetic cell isolation using nano-sized beads. Choose between various separation strategies for positive cell selection, sequential isolation, or cell depletion from any starting material, including peripheral blood mononuclear cells (PBMCs), whole blood, bone marrow, Leukopak® (StemExpress, LCC), LRSCs or buffy coats. MACS® Technology enables the magnetic separation of cell populations based on surface antigens. It is a fast and gentle method for the isolation of viable and functional cells by labeling epitopes with specific antibodies conjugated to superparamagnetic beads.

MACS® Technology—Magnetic cell isolation at a glance

• Flexible cell isolation options for your specific needs
• Column-based MACS® Technology using nano-sized beads
• Column-free technologies using micro-sized beads

With MACS Technology, you are sure to select the best.

MACS® MicroBeads – proven technology for basic research and clinical applications

MACS® MicroBeads are 50-nm superparamagnetic particles that are conjugated to highly specific antibodies against a particular cell surface antigen. Due to their small size, the beads do not activate cells. Furthermore, MACS MicroBeads do not have to be removed for any downstream application.

• MACS MicroBead Technology gives you the most flexible, most proven method for cell separation
• Minimal cell labeling with nano-sized MicroBeads ensures preservation of cellular integrity and characteristics
• Used in over 55,000 clinical cellular treatments to date

MACS Columns – maximal magnetic power for minimal cell labeling

At the heart of MACS® MicroBead Technology is the MACS Column, containing a matrix composed of ferromagnetic spheres covered with a cell-friendly coating. 
When the column is placed in a MACS Separator, the spheres amplify the magnetic field by 10,000-fold, thus inducing a strong magnetic force within the column. The magnetic field efficiently retains cells labeled with the small, nano-sized beads.

The spacious matrix inside the MACS Columns ensures that unlabeled cells can easily flow through while minimally labeled cells (fig. 4) are gently yet effectively retained (fig. 5). This minimizes stress on the cells and allows for efficient washing while preventing cell aggregation. 

MACS® Columns enable gentle flow of cells. No pressure, sticking, or compression.

The MACS® Technology advantage - Select the best by combining MACS® Columns and MicroBeads

Compared to column-free cell separation technologies, cell separation using MACS MicroBeads only requires minimal labeling of target cells. The benefits are no non-specific labeling, no cell activation and thereby no alteration of cell characteristics.

Cell isolation from single-cell suspensions and dissociated tissues

MACS® MicroBeads and MicroBead Kits

Straightforward positive selection of target cells based on specific markers
The strong magnetic field generated by the matrix in the MACS® Column allows for minimal labeling of target cells with nano-sized MicroBeads. This ensures that plenty of surface epitopes remain free for subsequent fluorescent staining and flow cytometry analysis. Moreover, low  labeling concentrations and the small size of MACS MicroBeads do not lead to activation of target cells (fig. 8).

• The least manipulative positive selection method
• Preservation of cell functionality due to optimal labeling
• Biodegradable: labeled cells are ready for downstream applications

MACS® Cell Isolation Kits

Depletion of non-target cells to obtain pure, truly untouched cells
MACS® Cell Isolation Kits contain a cocktail of titrated antibodies and MACS MicroBeads for indirect magnetic labeling (fig. 9). They are the preferred choice if binding of
antibodies to the target cells is not desired. Minimal labeling
of unwanted cells with MACS MicroBeads avoids nonspecific labeling of target cells, leaving the target cells truly untouched (fig. 10). In contrast, column-free methods based on nano-sized beads from other manufacturers require high concentrations of labeling reagents resulting in non-specific labeling of the target cell fraction.

• High purity and recovery rates
• Fully compatible with any downstream application
• No non-specific labeling of target cells
  
  
  
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Cell isolation directly from blood products

StraightFrom® Technology
Cell isolation directly from blood products without density gradient centrifugation
StraightFrom® MicroBeads allow magnetic isolation of various leukocyte subsets from different starting materials by positive selection. With these kits, isolation of leukocyte subsets has never been easier and quicker. 

• Start directly with whole blood, buffy coat, and leukocyte reduction system chamber (LRSC)
• The isolated target cells are immediately ready for any downstream application
• Simple protocol with only a few handling steps
  
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MACSxpress® Technology
With high speed to untouched target cells
MACSxpress® Technology enables the fastest large-scale isolation of untouched cells directly from whole blood – without the need for any centrifugation. Micro-sized MACSxpress Beads allow for minimal labeling to prevent non-specific labeling and activation of target cells. Nontarget cells are removed by immunomagnetic depletion. Simultaneously, erythrocytes are sedimented to yield target cells of exceptional purity.

• Go from whole blood to pure cells within 20 minutes
• Obtain untouched target cells directly from whole blood
• No density gradient centrifugation, erythrocyte lysis or cell counting required
  

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Magnetic cell separation strategies with MACS® Technology

The diversity of the MACS® Technology portfolio provides ultimate flexibility for cell separation procedures. Choose between various isolation strategies such as positive selection, depletion, untouched isolation, or sequential sorting of cell subsets to get the cells you really need.

• Positive cell selection for isolation of a particular cell type
• Untouched target cell isolation by depletion of unwanted cells
• Sequential separation for isolation of cell subsets
  If a certain desired cell type or subset does not express a single unique marker, it is not possible to isolate these cells in a single positive selection step. Instead a combination of two consecutive separations based on different markers can be applied.
  
  Depletion followed by positive selection
  This approach is useful if an important marker for the target cells is also expressed on a fraction of undesired cells. To enable positive selection of the target cells based on this marker, the fraction of undesired cells needs to be depleted first. To this end, the undesired cells are magnetically labeled via antigens distinct from that common marker. During separation, the labeled cells are retained in the column. The flow-through fraction contains the target cells. These cells can then be labeled with MACS MicroBeads for that marker, and the target cells are 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 consecutive positive selections
  MACS MultiSort MicroBeads enable cell isolation by two sequential positive selection steps based on two different markers. For the first positive selection, cells are labeled with MACS MultiSort MicroBeads specific for the first marker. After elution from the column, the cells are incubated with MultiSort Release Reagent, which enzymatically removes the MultiSort MicroBeads from the cells. For the second positive selection, the target cells are magnetically labeled with MACS MicroBeads directed against the second marker and isolated on the column.
  
  
  
  Two consecutive positive selections or positive selection followed by depletion
  
  REAlease Technology  allows for magnetic cell isolation by positive selection of target cells and subsequent removal of any beads and labels from the cells. The technology relies on recombinantly engineered antibody fragments instead of whole antibodies for cell surface labeling. Monomeric fragments have a low affinity for cell surface markers. However, when multimerized as part of the REAlease Complex, the fragments bind markers with high avidity. Anti-Biotin MicroBeads bound to the REAlease Complex facilitate isolation of the target cells based on MACS Columns and Separators (see figure). When the column is placed in the magnetic field of the separator, unlabeled cells flow through, whereas the target cells are retained on the column. Subsequently, labeled target cells are eluted from the column using REAlease Bead Release Reagent, which removes Anti-Biotin MicroBeads from the cells.
  Following elution, there are two options to isolate subsets from this population:
  I) Disruption of the REAlease Complex by adding REAlease Release Reagent leads to monomerization and spontaneous dissociation of the antibody fragments from the cell surface. Cells are then label free and suitable for magnetic re-labeling with REAlease Technology.
  II) Alternatively, eluted cells can be directly labeled with MACS MicroBeads.
  
  • Bead-free cells: suited for second round of magnetic labeling
  • Label-free cells: the epitope of a marker becomes completely available again
  • Recombinantly produced: lot-to-lot consistency allows for reproducible results
  
  
  Direct magnetic labeling
  
  Direct labeling with MACS MicroBeads is the fastest way of magnetic labeling. MACS MicroBeads specifically bind to antigens on the cell surface. Only one incubation step is required. Direct magnetic labeling requires a minimal number of washing steps and therefore minimizes cell loss.
  Highly specific cell separation reagents for direct labeling of numerous cell types with MACS MicroBeads are available for human, mouse, rat, and non-human primate cells.
  
  Indirect magnetic labeling
  Combination of primary antibodies and MicroBeads
  
  

  Indirect magnetic labeling using primary antibodies and MicroBeads is based on a two-step procedure. First, the cells are labeled with a primary antibody directed against a cell surface marker. Subsequently, the cells are magnetically labeled with a MACS MicroBead, which either binds directly to the primary antibody or to a molecule that is conjugated to the primary antibody. Conjugated molecules include biotin and fluorochromes. Accordingly, magnetic labeling is achieved with Anti-Immunoglobulin MicroBeads, Anti-Biotin MicroBeads, or Anti-Fluorochrome MicroBeads.

  This indirect labeling strategy is useful for isolating untouched target cells. In this case, the unwanted cell types are incubated concurrently with a cocktail of primary antibodies conjugated to biotin, for example. Subsequently, cells are labeled with Anti-Biotin MicroBeads.

  REAlease Technology

  

  REAlease Technology is an indirect cell labeling method for positive selection of target cells. The REAlease Biotin Complex binds to the target cells. Labeling of the REAlease Biotin Complex with Anti-Biotin MicroBeads allows for magnetic isolation of these cells. Following cell separation, both MicroBeads and REAlease Biotin Complex can be gently removed, leaving the cells bead-and label-free. Enriched cells are then suitable for magnetic re-labeling and any application where label-free cells are essential.

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