Recombinant Human KEAP1/CUL3/NEDD8/RBX1 Complex Protein, CF

R&D Systems | Catalog # E3-701

Discontinued.
R&D Systems
Discontinued Product
E3-701 has been discontinued. View all KEAP1/CUL3/NEDD8/RBX1 products.

Key Product Details

Source

Sf 21 (baculovirus)

Accession Number

Q14145, Q13618, Q15843, P62877

Applications

Bioactivity
View all KEAP1/CUL3/NEDD8/RBX1 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human KEAP1/CUL3/NEDD8/RBX1 protein

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Predicted Molecular Mass

70 kDa (KEAP1), 101 kDa (neddylated CUL3), 12 kDa (RBX1)

Activity

Typical protein concentration for use in vitro will depend on experimental conditions.  For in vitro ubiquitination reactions, we suggest an initial KEAP1/CUL3(NEDD8)/RBX1 concentration between 10-50 nM.

Formulation, Preparation, and Storage

E3-701
Formulation X mg/ml (X μM) in 50 mM HEPES pH 7.5, 200 mM NaCl, 10% (v/v) Glycerol, 1 mM DTT
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 48 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: KEAP1/CUL3/NEDD8/RBX1

Kelch-like ECH-associated protein (KEAP1) is the substrate recognition subunit of a BCR (BTB-CUL3-RBX) E3 ubiquitin ligase complex.  KEAP1 contains an N-terminal BB Domain that mediates interaction with CUL3, a central BACK Domain that plays a role in protein-protein interactions, and six C-terminal Kelch Domains that function in substrate recognition.  In the BCR complex, CUL3 serves as a scaffold that organizes one or more BTB (BR-C, Ttk and Bab, also known as a POZ Domain) substrate recognition subunits with the RBX subunit and contributes to catalysis through positioning of the substrate and an E2 ubiquitin-conjugating enzyme.  KEAP1 is an important sensor of oxidative and electrophilic stress.  Normally the KEAP1/CUL3/RBX1 complex ubiquitinates and causes degradation of NRF2, a transcription factor regulating expression of many cytoprotective genes.  Oxidative stress results in the inability of KEAP1 to ubiquitinate NRF2, leading to the latter proteins accumulation and subsequent upregulation of genes involved in cellular oxidative stress responses.  This recombinant complex consists of an N-terminal 10-His tagged Cullin-3 (UniProt Q13618), untagged RBX1 (UniProt P62877) and untagged KEAP1 (UniProt Q14145-1).  Cullin-3 is neddylated (Q15843) in this complex.

References

  1. Eggler, A.L. et al. (2009) Biochem. J.  doi:10.1042/BJ20090471
  2. Eggler, A.L. et al. (2005) Proc. Natl. Acad. Sci.  doi: 10.1073/pnas.0502402102
  3. Mills, E.L. et al. (2018) Nature   doi: 10.1038/nature25986
  4. Wei, J. et al. (2021) JACS   doi: 10.1021/jacs.1c04841

Long Name

KEAP1/CUL3/NEDD8/RBX1

UniProt

Q14145, Q13618, Q15843, P62877

Product Documents for Recombinant Human KEAP1/CUL3/NEDD8/RBX1 Complex Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human KEAP1/CUL3/NEDD8/RBX1 Complex Protein, CF

For research use only

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