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KLF4 Expression Vectors

The APX/APEX kits have been discontinued effective June 30, 2010. Please see the QIAgenes Expression Kits.

Adjustable KLF4 Plasmid Adjustable KLF4 Lentivirus Constitutive KLF4 Plasmid Constitutive KLF4 Lentivirus
 
APEX KLF4 Destabilization Domain Expression Plasmid
Human KLF4-DD Plasmid: DB7-103A
Mouse KLF4-DD Plasmid:
DB2-603A

Human KLF4-DD Plasmid
Mouse KLF4-DD Plasmid
TThe APEX KLF4 Destabilization Domain Expression Plasmid is a ready-to-transfect plasmid that encodes either a human or mouse DD-fused KLF4 protein. The open reading frame that codes for the fusion protein is under the control of the constitutively active CMV promoter. The amount of DD-fused KLF4 protein is proportional to the amount of Centry DD-protein Stabilization Reagent present. Without the Centry reagent, the unstable DD-fusion protein is recognized by the proteasome and rapidly degraded. Therefore, the expression of the KLF4 protein is adjustable based on the concentration of the Centry reagent.

The KLF4-DD expression plasmid can easily and rapidly:

  • Express KLF4 protein in a dose-dependent manner under the control of Centry
  • Generate iPSCs when expressed in combination with other APEX vectors
  • Generate stable Oct-DD expressing cells

See the complete list of APEX Vectors.

See the parental expression vector or control vectors.

User Manual 

 

Kit Components How It Works Manual & Resources Related products
 
Component Specification Concentration* (total volume)
KLF4-DD Plasmid  An expression plasmid for either Human or Mouse KLF4 fused with a Destabilization Domain under the control of a CMV promoter.  10 micrograms
 

Kit Components How It Works Manual & Resources Related Products
 
  • APEX Technology Overview

The APEX DD-containing vectors encode proteins that have adjustable expression at the protein level. The DD-fusion protein is transcriptionally under the control of a CMV promoter (1). In the presence of Centry the protein is stabilized, because Centry binds to the DD domain allowing the fusion protein to accumulate in the cell in a dose-dependent manner (2). In the absence of Centry or the removal of Centry from the media, the proteasome degrades the DD-fusion protein (3).

  • Sample APEX Expression Data

Adjust protein expression by changing the media.

HEK293 cells were transfected with the DD-fused GFP. At the beginning of the experiment, media with 500 nM Centry was added to the cells to stabilize the GFP molecule. After 24 hours, the media was replaced with standard growth media. Fluorescent micrographs were taken at the time points indicated.

APEX System Delivers Dose-Dependent Protein Expression.

HEK293 cells were stably transfected with DD-fused GFP. Cells were treated with increasing concentrations of APEX Centry at the beginning of the experiment. APEX Centry was then removed after overnight incubation by removing the media and adding fresh growth media. Fluorescent Activity is expressed in arbitrary units and was monitored throughout the experiment with the use of a fluorometer.

APEX Expression Constructs Yield Functional Proteins.

HEK293 cells were transiently transfected with Cignal™ KLF4 luciferase reporter (CCS-4036L) and the APEX DD-fused KLF4. A renilla luciferase expression plasmid was used as a transfection efficiency control. The media was changed after transfection to either growth media or media containing 500 nM Centry. Cells were lysed after 24 hours and assayed for luciferase activity. Relative luciferase activity is shown as the mean ( S.D.) of three independent experiments.

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Kit Components How It Works Manual & Resources Related Products
 
User Manuals APEX System User Manual (PDF)
Flyer APEX System (PDF)
Vector Map
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Kit Components How It Works Manual & Resources Related Products
 
APEX Centry  Small molecule for directly regulating APEX protein expression
SureFECT Transfection  Achieve high transfection efficiencies and minimal cytotoxicity
Pathway reporters  Complete list of 38 signaling pathway reporters
10-Pathway Arrays  Simultaneously measure the activities of ten (10) signaling pathways in a single experiment

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