MACSelectâ„¢ 4 System
The MACSelectâ„¢ 4 System uses the truncated human CD4 molecule as a marker to select transfected cells. It can be used in virtually all CD4-negative cell lines and primary cells.* pMACS 4-IRES.II enables the bicistronic expression of the gene-of-interest and the truncated CD4 molecule: with the IRES element (internal ribosomal entry site), the gene-of-interest is translated from the same mRNA as the selection marker CD4. The pMACS 4.1 vector encoding the truncated CD4 surface marker is used for co-transfection in combination with a gene-of-interest expression vector. The MACSelect 4 Transfected Cell Selection Kit provides the vectors, MACSelect MicroBeads, and fluorochrome-conjugated antibodies, including control vectors and antibodies required to set up and establish the MACSelect System. All kit components are also available separately. * The CD4 molecule is sensitive to trypsin treatment and should, therefore, not be used in cell lines which need to be harvested with trypsin. Columns For positive selection: MS, LS, or autoMACSâ„¢ Columns. Disclaimer The CMV promoter is covered under U.S. patents 5,168,062 and 5,385,839 and its use is permitted for research purposes only. Any other use of the CMV promoter requires a licence from the University of Iowa Research Foundation, 214 Technology Innovation Center, Iowa City, IA 52242. Further information Plasmid transfection protocols [PDF; 139,8 KB] DNA sequence pMACS 4-IRES.II (Text Document, PC) [Raw Text File; 5,7 KB] DNA sequence pMACS 4-IRES.II (RTF Document, MAC) [RichTextFile (RTF); 10,5 KB] vector map pMACS 14.1 (Single Cutting Restriction Sites) [PDF; 20,3 KB] vector map pMACS 14.1 (Twice Cutting Restriction Sites) [PDF; 22,3 KB] vector map pMACS 4.1 (Single Cutting Restriction Sites) [PDF; 22 KB] vector map pMACS 4.1 (Twice Cutting Restriction Sites) [PDF; 23,7 KB] DNA sequence pMACS 4.1 (Text Document, PC) [Raw Text File; 4,4 KB]		Figure 1 The pMACS 4-IRES.II vector. * The BsmB I site is compatible with Hind III digested fragments. Figure 2 The pMACS 4.1 vector. Figure 3 Transfection of CHO cells with pMACS 4-IRES.II, magnetic labeling with MACSelect 4 MicroBeads and separation. FITC staining of transfected cells (CD4-FITC/MACSelect Control FITC), blue nuclear counterstain (Toto 3, Molecular Probes). A: Less than 4% transfected cells after transfection B: More than 90% transfected cells after MACSelect enrichment Figure 4 MACSelect transfected cell enrichment