Enabling Protein Degradation Drug Discovery

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  • Name
    Catalogue Number
    Size
    Price
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  • Name:
    HEK293 cell lysate (IL-1 stimulated)
    Catalogue Number:
    66-3001-010
    Size:
    10 mg
    Price:
    £325
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  • Species
    Human
  • Source
    IL-1-stimulated embryonic kidney (HEK293) cells
  • Quantity
    10 mg
  • Storage
    -70°C
  • Concentration
    20mg/ml
  • Formulation
    50mM Tris/HCl pH7.5, 1mM EGTA, 1mM EDTA, 1% Triton X-100, 1mM sodium orthovanadate, 50mM sodium fluoride, 5mM sodium pyrophosphate, 10mM sodium β-glycerophosphate, 270mM sucrose, 1mM PMSF, 1mM benzamidine, 50mM iodoacetamide.
  • Molecular Weight
    n/a
  • Stability
    12 months at -70°C; aliquot as required
  • Protein Sequence
    n/a
  • QA; Protein Identification
    n/a
  • QA; Activity
    Identification of Polyubiquitin Chains in Lysate: A Western blot of IL-1 stimulated HEK293 cell lysate (lane 2) and non-stimulated HEK293 cell lysate (lane 3) probed with an anti-FK2 antibody indicates that both of the lysates are abundant with ubiquitylated proteins and possibly free ubiquitin chains. Polyubiquitin (Ub2-7) (K63-linked) was included as a positive control in this experiment (lane 1).

HEK293 Cell Lysate (IL-1 stimulated) may be used to demonstrate the ability of  ubiquitin binding domain proteins (Optineurin and NEMO) to capture polyubiquitylated proteins including IL-1 receptor-associated kinase 1 (IRAK1) (Windheim et al. 2008).

Reference:Windheim M, Stafford M, Peggie M, Cohen P (2008) Interleukin-1 (IL-1) induces the Lys63-linked polyubiquitination of IL-1 receptor-associated kinase 1 to facilitate NEMO binding and the activation of IkappaBalpha kinase. Mol Cell Biol 28, 1783-91.