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human pirh2-(187-261)
(Plasmid #24883)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 24883 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pGEX-2TKm
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Pirh2
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    222
  • Mutation
    Contains aa 187-261.
  • Entrez Gene
    RCHY1 (a.k.a. ARNIP, CHIMP, PIRH2, PRO1996, RNF199, ZCHY, ZNF363)
  • Tag / Fusion Protein
    • GST (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer pGEX5'
  • 3′ sequencing primer pGEX3'
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

E. coli strain for expression: BL21DE3

Expression conditions: Pirh2 proteins need Zn2+ to fold properly, so 50 uM of ZnCl2 is added to the growth media and purification buffers.

Induction conditions: 0.5mM-1.0mM IPTG at RT for 3.5-4 hours.

For pGEX-2TKm vector, the kinase site RRASV is removed and NdeI site is inserted at 5' of BamHI.

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:

    human pirh2-(187-261) was a gift from Cheryl Arrowsmith (Addgene plasmid # 24883 ; http://n2t.net/addgene:24883 ; RRID:Addgene_24883)

  • For your References section:

    Molecular basis of Pirh2-mediated p53 ubiquitylation. Sheng Y, Laister RC, Lemak A, Wu B, Tai E, Duan S, Lukin J, Sunnerhagen M, Srisailam S, Karra M, Benchimol S, Arrowsmith CH. Nat Struct Mol Biol. 2008 Dec . 15(12):1334-42. 10.1038/nsmb.1521 PubMed 19043414
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