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V51 pIRESpuro-GLUE (pGLUE)
(Plasmid #15100)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 15100 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pIRESpuro-GLUE
  • Backbone manufacturer
    Moon Lab
  • Backbone size (bp) 5500
  • Vector type
    Mammalian Expression
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    None
  • Tags / Fusion Proteins
    • streptag (N terminal on backbone)
    • HA (N terminal on backbone)
    • CBP tag (N terminal on backbone)

Cloning Information

Resource Information

Depositor Comments

To generate the pGLUE tandem-affinity purification plasmid, the pIRES–puro (Clontech, Mountain View, CA) was used as the backbone vector and the dual affinity tag was constructed using PCR.

The pGLUE expression vector was designed to have the streptavidin-(streptag) and calmodulin-(CBP) binding affinity tags placed in tandem at the N-terminus of the bait protein. An HA epitope is also present to facilitate protein detection. The first round of affinity purification is performed using a streptavidin resin, followed by the specific elution with biotin and the protease TEV. The second round is done using the calcium dependent binding to the calmodulin resin and specific elution by chelation using EGTA.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    V51 pIRESpuro-GLUE (pGLUE) was a gift from Randall Moon (Addgene plasmid # 15100 ; http://n2t.net/addgene:15100 ; RRID:Addgene_15100)
  • For your References section:

    The KLHL12-Cullin-3 ubiquitin ligase negatively regulates the Wnt-beta-catenin pathway by targeting Dishevelled for degradation. Angers S, Thorpe CJ, Biechele TL, Goldenberg SJ, Zheng N, MacCoss MJ, Moon RT. Nat Cell Biol. 2006 Apr . 8(4):348-57. 10.1038/ncb1381 PubMed 16547521