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| Questions & answers |
| What are the differences between the three MACSelect™ Systems? |
All three systems work according to the same principle: by co-expression of a truncated surface marker, it is possible to magnetically label and separate the transfected cells — without any unwanted signaling or antibiotic effects on the cell. The gentle MACS® Separation with the extremely small (approx. 50 nm diameter) MicroBeads has no effect on cell function and viability.
The differences between the MACSelect™ Systems are the surface markers and the pMACS vectors that encode them.
With a choice of three different surface markers, CD4, H2Kk, and LNGFR*, virtually any cell type can be enriched via MACSelect™ System. A suitable MACSelect surface marker must not naturally be expressed on the cells to be transfected.
For straightaway use of the MACSelect System, cells can be co-transfected with a pMACS vector and an expression vector containing the gene-of-interest. Alternatively, the gene-of-interest can be cloned into a pMACS vector for single-vector transfection. Please see the catalog or visit www.MACSmolecular.com for details.
MACSelect Kits contain MACSelect MicroBeads, all necessary vectors for either cloning or co-transfection, and antibody controls. All kit components are also available as single reagents.
*CD4 is naturally expressed on T helper cells, monocytes and dendritic cells. MACSelect 4 should not be used for such cell types of human origin. CD4 is trypsin sensitive. H-2Kk expression is restricted to some rarely used mouse strains like AKR/J and CBA/Ca. MACSelect Kk should not be used for such murine cell types. H-2Kk requires co-expression of ß-2-microglobulin. LNGFR is expressed in the CNS and PNS on autonomic and sensory neurons and glial cells, on bone marrow fibroblasts, follicular dendritic cells, and some mesenchymal cells. MACSelect LNGFR should not be used for such cell types of human origin. |
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| Can the MACSelect™ System only be used for enrichment of transiently transfected cells? |
No, you can use it as well for the establishment of stable cell lines.
The MACSelect™ System can be used to enrich transiently transfected cells by magnetic cell selection a few hours after transfection. Additionally, magnetic cell selection can be used to establish a stable cell line without the use of any antibiotic. For this purpose, the enriched cells will be cultured for a few days before a second round of separation is performed. During this cultivation period the MACS® MicroBeads are biodegraded. Only stably transfected cells, which have integrated the plasmid into their genome, will further express the truncated surface marker and can be magnetically labeled and separated. |
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