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CherryPicker System

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Capture only those cells with promoter activity
Use the promoterless CherryPicker systems (CherryPicker Cell Capture System and Lenti-X™ CherryPicker Cell Capture System) to monitor the activation of a promoter of interest, and capture those cells containing the active promoter. Insertion of a functional promoter into the promoterless reporter vector, pCherryPicker1, causes a red fluorescent protein (CherryPicker) to be displayed on the surface of mammalian cells, which are then easily captured by a CherryPicker-specific antibody bound to IgG-coated magnetic beads.
Capture only those cells expressing your protein
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 population



Figure 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 interest into 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 population was 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-specific Erk1/2 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.
Applications
Promoterless Systems:
- Monitor the activation of a promoter of interest
- Capture and analyze cells in which a promoter of interest is active
- Culture the captured cells as a more homogeneous population

Bicistronic (IRES) Systems:
- Monitor cells expressing a protein of interest
- Capture and analyze cells expressing the protein of interest
- Culture the captured cells as a more homogeneous population
Components
CherryPicker Assay Kit
CherryPicker Antibody
Wash Buffer
IgG-Coated Magnetic Capture Beads

CherryPicker Cell Capture System
pCherryPicker1 and pCherryPicker Control Vectors
CherryPicker Assay Kit

CherryPicker Cell Capture System (IRES)
pCherryPicker2 and pCherryPicker Control Vectors
CherryPicker Assay Kit

Lenti-X CherryPicker Cell Capture System
pLVX-CherryPicker1 and pLVX-CherryPicker Control Vectors
CherryPicker Assay Kit

Lenti-X CherryPicker Cell Capture System (IRES)
pLVX-CherryPicker2 and pLVX-CherryPicker Control Vectors
CherryPicker Assay Kit
Storage Conditions
CherryPicker Vectors & Antibody: -20°C
Wash Buffer & Magnetic Capture Beads: 4°C

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