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Expression Vectors for Pierce qIP Kits


Luciferase (Tluc) and affinity tag vectors for qIP systems.

Pierce qIP Expression Vectors

These Thermo Scientific pCMV Vectors include epitope-tagged (HA or c-Myc) and Tluc-tagged (TurboLuc Luciferase) multiple cloning site mammalian expression vectors for use in Pierce quantitative IP (qIP) assays.

Pierce qIP Assays are designed to measure the interaction of two proteins (X and Y) that are transiently co-expressed in mammalian cells as epitope-tagged and luciferase-tagged fusions, respectively. The specific luciferase required is Thermo Scientific TurboLuc (Tluc) Luciferase, which is especially small and bright. Protein interactions are quantified by measuring Tluc Luciferase activity following immunoprecipitation (IP) of epitope-tagged proteins with anti-epitope agarose or magnetic beads. As such, the assay system requires that the genes for proteins-of-interest X and Y be cloned as fusions with epitope (e.g., HA or c-Myc) and Tluc tags, respectively. Several vectors, all based on a common pCMV-MCS map, are available for this purpose. These optimized MCS vectors include varieties with tags at the N- or C-terminus. Also available are ready-to-use, positive and negative control vectors containing genes for BAD, RFP or Bcl-xL proteins.

Highlights:

  • CMV (Cytomegalovirus) promoter for constitutive expression of proteins in mammalian cells
  • Shared multiple cloning site (MCS) in all vectors provides convenience and versatility for transfer of cDNA gene from one plasmid to another
  • Tluc tagged vectors use the small and bright TurboLuc Luciferase gene, providing high expression in mammalian systems and the sensitivity required for quantitative IP
  • HA epitope tag (YPYDVPDYA) and Myc epitope tag (EQKLISEEDL) varieties available
  • N-terminal and C-terminal tagged vectors offered for each tag, enabling optimization of protein function for protein:protein interaction
  • Kanamycin/neomycin marker for drug selection in both bacterial and mammalian cells
  • Complete cloning and subcloning guide available (Tech Tip #77)

Product Details:

All Pierce qIP Vectors contain the same multiple cloning site (MCS) sequences. Therefore, once a gene of interest is successfully cloned into one qIP vector with the proper reading frame, the gene can be moved to another qIP vector without additional PCR amplification. This feature provides the convenient and flexible option to subclone a gene from an N-terminal tag vector to C-terminal tag vector.

BAD and Bcl-xL proteins interact strongly as a protein interaction pair. Thus, pairing an epitope-tagged BAD vector with a Tluc-tagged Bcl-xL vector forms a positive control for Pierce qIP Assays. Red fluorescent protein (RFP) does not interact with either BAD or Bcl-xL. Therefore, pairing an epitope-tagged RFP vector with Tluc-tagged Bcl-xL vector provides a negative control for Pierce qIP Assays. The prepared epitope-tagged control vectors (BAD, RFP) that are included in the Pierce qIP Kits are cloned into the nominal MCS expression vectors using NotI and BglII restriction sites. As such, these control vectors can be used as expression vectors for cloning or subcloning a gene of interest.

Download the complete sequence (.txt) and printable data sheet (.pdf) of each vector. Vector names consists of the following parts: promoter type (CMV), gene name (or MCS), tag position (N or C) and tag name (HA, Myc or Tluc). For example, pCMV BAD C-HA expresses BAD with C-terminal HA tag driven by the CMV promoter.
Vector Sequence File Data Sheet
pCMV MCS N-HA Seq-82017-pCMV-MCS-N-HA.txt CofA-82017-specs.pdf
pCMV MCS C-HA Seq-82018-pCMV-MCS-C-HA.txt CofA-82018-specs.pdf
pCMV MCS N-Myc Seq-82019-pCMV-MCS-N-Myc.txt CofA-82019-specs.pdf
pCMV MCS C-Myc Seq-82020-pCMV-MCS-C-Myc.txt CofA-82020-specs.pdf
pCMV MCS N-Tluc Seq-82023-pCMV-MCS-N-Tluc.txt CofA-82023-specs.pdf
pCMV MCS C-Tluc Seq-82024-pCMV-MCS-C-Tluc.txt CofA-82024-specs.pdf
pCMV RFP C-HA Seq-82025-pCMV-RFP-C-HA.txt CofA-82025-specs.pdf
pCMV RFP C-Myc Seq-82026-pCMV-RFP-C-Myc.txt CofA-82026-specs.pdf
pCMV BAD C-HA Seq-82028-pCMV-BAD-C-HA.txt CofA-82028-specs.pdf
pCMV BAD C-Myc Seq-82029-pCMV-BAD-C-Myc.txt CofA-82029-specs.pdf
pCMV Bcl-xL N-Tluc Seq-82031-pCMV-Bcl-xL-N-Tluc.txt CofA-82031-specs.pdf

These vectors are subject to a limited use label license.

General References:

  • Green, M.R., and Sambrook, J. (2012). Molecular Cloning: A Laboratory Manual (Fourth Edition). Cold Spring Harbor Laboratory Press.
  • Kozak, M. (1990). Downstream secondary structure facilitates recognition of initiator codons by eukaryotic ribosomes. Proc. Natl. Acad. Sci. USA 87: 8301-5.

Related Resources:

Overview of Pierce qIP Protein Interaction Kits
Tech Tip #77: Cloning guide for Pierce qIP Protein Interaction Kits

Related Products:

Pierce qIP Protein Interaction Kits
Pierce qIP Buffers and Assay Reagents


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
82017 pCMV MCS N-HA
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82017 pCMV MCS N-HA Certificate of Analysis for product #82017 pCMV MCS N-HA $75.00

Add to cart
82018 pCMV MCS C-HA
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82018 pCMV MCS C-HA Certificate of Analysis for product #82018 pCMV MCS C-HA $75.00

Add to cart
82019 pCMV MCS N-Myc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82019 pCMV MCS N-Myc Certificate of Analysis for product #82019 pCMV MCS N-Myc $75.00

Add to cart
82020 pCMV MCS C-Myc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82020 pCMV MCS C-Myc Certificate of Analysis for product #82020 pCMV MCS C-Myc $75.00

Add to cart
82023 pCMV MCS N-Tluc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82023 pCMV MCS N-Tluc Certificate of Analysis for product #82023 pCMV MCS N-Tluc $75.00

Add to cart
82024 pCMV MCS C-Tluc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82024 pCMV MCS C-Tluc Certificate of Analysis for product #82024 pCMV MCS C-Tluc $75.00

Add to cart
82025 pCMV RFP C-HA
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82025 pCMV RFP C-HA Certificate of Analysis for product #82025 pCMV RFP C-HA $75.00

Add to cart
82026 pCMV RFP C-Myc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82026 pCMV RFP C-Myc Certificate of Analysis for product #82026 pCMV RFP C-Myc $75.00

Add to cart
82028 pCMV BAD C-HA
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82028 pCMV BAD C-HA Certificate of Analysis for product #82028 pCMV BAD C-HA $75.00

Add to cart
82029 pCMV BAD C-Myc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82029 pCMV BAD C-Myc Certificate of Analysis for product #82029 pCMV BAD C-Myc $75.00

Add to cart
82031 pCMV Bcl-xL N-Tluc
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #82031 pCMV Bcl-xL N-Tluc Certificate of Analysis for product #82031 pCMV Bcl-xL N-Tluc $75.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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