□ Capture only those cells with promoter activity

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□ Capture only those cells expressing your protein

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Use the bicistronic CherryPicker systems [CherryPicker Cell Capture System (IRES) and Lenti-X CherryPicker Cell Capture System (IRES)] to identify, monitor, and capture mammalian cells expressing your protein of interest. IRES technology results in simultaneous expression of a protein of interest and a membrane-targeted red fluorescent protein (CherryPicker) from the same transcript. Cells expressing your protein of interest must also express CherryPicker, and can be captured on magnetic beads via a CherryPicker-specific antibody. Coexpression of your protein of interest and CherryPicker allows you to easily:1.Monitor cells expressing the protein of interest 2.Capture and analyze cells expressing the protein of interest 3.Culture the captured cells as a homogeneous populationFigure 1. Select cells for promoter activity or protein expression, using the CherryPicker Systems. Panel A. Use the promoterless systems to capture cells with promoter activity. Inserting your promoter of interest into the promoterless reporter vector causes a red fluorescent protein (CherryPicker)to be displayed on the surface of mammalian cells, which are then easily captured on magnetic beads via a CherryPicker-specific antibody.Panel B. Use the bicistronic (IRES) systems to capture cells expressing your protein of interest. Insert the gene for your protein of interestinto the IRES reporter vector. The protein will be expressed simultaneously with a membrane-targeted red fluorescent protein (CherryPicker)from the same transcript. Cells expressing your protein of interest can then be captured on magnetic beads via a CherryPicker-specific antibody. Figure 2. Highly efficient enrichment of CherryPicker cells from a mixed population. Two populations of cells were created, one expressing AcGFP1 and the other expressing CherryPicker. The two populations were mixed in a ratio of > 6:1 AcGFP1:CherryPicker cells. The mixed populationwas then enriched for CherryPicker-expressing cells using the CherryPicker Assay Kit. After enrichment, the ratio was > 1:4 AcGFP1:CherryPicker cells.The distribution of cells before and after capture was determined by flow cytometry.Figure 3. Easily select cells expressing a protein of interest using the Bicistronic (IRES) CherrryPicker Systems. HEK-293 cells were transiently transfected with a pCherryPicker2-MEK-CA construct. A HEK-293 cell population containing 10% transfected cells was divided into two equal fractions. One fraction of cells was lysed without any further treatment (middle lanes). The second fraction of cells was enriched for MEK-CA expressing cells using the CherryPicker magnetic bead system prior to lysis (right lane). Both lysates were analyzed by western blot using the anti-phospho-specificErk1/2 antibody. Lane1: Lysate of 35 x103 non-transfected HEK293 cells. Lanes 2-4: Lysates equivalent of 17.5, 35, and 52.5 x103 cells, from the fraction that was lysed without enrichment. Lane 5: Lysate of captured cells from the fraction that was enriched using the magnetic bead-based CherryPicker kit. To allow for normalization, the western blot was also tested using an antibody against the ubiquitously expressed GAPDH.

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□ Applications

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□ Components

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□ Storage Conditions

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