pCMV MCS N-Myc Vector for Quantitative IP (qIP) Assays

Catalog number: 82019

Thermo Scientific™  Related applications: Protein Assays and Analysis | Protein Expression

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Description

The Thermo Scientific pCMV MCS N-Myc vector is an epitope-tagged multiple cloning site mammalian expression vector for use in Pierce quantitative IP (qIP) assays.

Features of the pCMV MCS N-Myc vector:

• CMV (Cytomegalovirus) promoter for constitutive expression of proteins in mammalian cells
• Shared multiple cloning site (MCS) provides convenience and versatility for transfer of cDNA gene from one plasmid to another
• N-terminal Myc epitope tag (EQKLISEEDL)
• Kanamycin/neomycin marker for drug selection in both bacterial and mammalian cells
• Complete cloning and subcloning guide available ()

About the Pierce qIP Assay
Pierce qIP Assays are designed to measure the interaction between 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.

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.

These vectors are subject to a limited use label license.

More Product Data
Pierce qIP Assay: a novel, HTS-compatible protein-protein interaction assay system

Related Products
pCMV MCS C-Myc Vector for Quantitative IP (qIP) Assays
pCMV MCS N-Tluc Vector for Quantitative IP (qIP) Assays
pCMV MCS C-Tluc Vector for Quantitative IP (qIP) Assays
pCMV RFP C-Myc Vector for Quantitative IP (qIP) Assays
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: CMV
Key Function: Protein-Protein Interaction Studies
Protein Tag or Fusion: c-Myc Epitope Tag
Delivery Method: Transfection
Cloning Method: Restriction Enzyme ⁄ MCS
Selection Agent (Eukaryotic): Neomycin⁄Kanamycin
Product Size: 10 µg

Contents & storage

Store between -70°C and -20°C.

Documents

Manuals & protocols