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

His10 In the process of being discontinued
Catalog # Availability Size / Price Qty
E3-701-025
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human KEAP1/CUL3/NEDD8/RBX1 Complex Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
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.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human KEAP1/CUL3/NEDD8/RBX1 protein
Predicted Molecular Mass
70 kDa (KEAP1), 101 kDa (neddylated CUL3), 12 kDa (RBX1)

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E3-701

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.
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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
Alternate Names
KEAP1/CUL3/NEDD8/RBX1

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