Enabling Protein Degradation Drug Discovery

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  • Name
    Catalogue Number
    Size
    Price
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  • Name:
    HEK293 cell lysate
    Catalogue Number:
    66-3002-010
    Size:
    10 mg
    Price:
    £250
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  • Species
    Human
  • Source
    Human Embryonic kidney
  • 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 may be used to demonstrate the ability of ubiquitin binding domain proteins (Op tineurin and NEMO) to capture polyubiq uitylated 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.