HEK293 cell lysate


Catalogue Number
66-3002-010
Product Size
10 mg
Price £
£250
Accession Number
n/a
Residues Expressed
n/a
Certificate of Analysis Size
10 mg
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).


Background

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).


References

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.