KEAP1 Protein, Human (sf9)

KEAP1 in the BCR E3 ubiquitin ligase complex complex regulates cellular responses to oxidative stress by ubiquitinating NFE2L2/NRF2. As an oxidative stress sensor, KEAP1 normally promotes NFE2L2/NRF2 degradation. KEAP1 Protein, Human (sf9) is the recombinant human-derived KEAP1 protein, expressed by Sf9 insect cells , with tag free. The total length of KEAP1 Protein, Human (sf9) is 623 a.a., with molecular weight of ~64 kDa.

KEAP1, as the substrate-specific adapter within the BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complex, intricately regulates the cellular response to oxidative stress by orchestrating the ubiquitination of NFE2L2/NRF2. Serving as a crucial sensor for oxidative and electrophilic stress, KEAP1, under normal conditions, facilitates the ubiquitination and subsequent degradation of NFE2L2/NRF2, a transcription factor essential for the expression of numerous cytoprotective genes. When confronted with oxidative stress, distinct electrophile metabolites induce non-enzymatic covalent modifications on highly reactive cysteine residues in KEAP1, effectively dampening the ubiquitin ligase activity of the BCR(KEAP1) complex. This disruption promotes the nuclear accumulation of NFE2L2/NRF2 and triggers the expression of phase II detoxifying enzymes. Furthermore, selective autophagy leads to the sequestration of KEAP1 in inclusion bodies through its interaction with SQSTM1/p62, resulting in the inactivation of the BCR(KEAP1) complex and the activation of NFE2L2/NRF2. Notably, the BCR(KEAP1) complex extends its ubiquitin ligase activity to substrates like SQSTM1/p62, BPTF, and PGAM5, modulating their degradation via the proteasome. The ubiquitin ligase activity of the BCR(KEAP1) complex faces inhibition in response to oxidative stress and electrophile metabolites such as sulforaphane, as these metabolites react with reactive cysteine residues in KEAP1, leading to the non-enzymatic covalent modifications that incapacitate the complex. Moreover, selective autophagy contributes to the inactivation of the BCR(KEAP1) complex through the interaction between KEAP1 and SQSTM1/p62, promoting the sequestration of the complex in inclusion bodies and facilitating its degradation.

Measured by its binding ability in a functional ELISA.

Human

Sf9 insect cells

Tag Free

Q14145 (Q2-C624)

9817  [NCBI]

QPDPRPSGAGACCRFLPLQSQCPEGAGDAVMYASTECKAEVTPSQHGNRTFSYTLEDHTKQAFGIMNELRLSQQLCDVTLQVKYQDAPAAQFMAHKVVLASSSPVFKAMFTNGLREQGMEVVSIEGIHPKVMERLIEFAYTASISMGEKCVLHVMNGAVMYQIDSVVRACSDFLVQQLDPSNAIGIANFAEQIGCVELHQRAREYIYMHFGEVAKQEEFFNLSHCQLVTLISRDDLNVRCESEVFHACINWVKYDCEQRRFYVQALLRAVRCHSLTPNFLQMQLQKCEILQSDSRCKDYLVKIFEELTLHKPTQVMPCRAPKVGRLIYTAGGYFRQSLSYLEAYNPSDGTWLRLADLQVPRSGLAGCVVGGLLYAVGGRNNSPDGNTDSSALDCYNPMTNQWSPCAPMSVPRNRIGVGVIDGHIYAVGGSHGCIHHNSVERYEPERDEWHLVAPMLTRRIGVGVAVLNRLLYAVGGFDGTNRLNSAECYYPERNEWRMITAMNTIRSGAGVCVLHNCIYAAGGYDGQDQLNSVERYDVETETWTFVAPMKHRRSALGITVHQGRIYVLGGYDGHTFLDSVECYDPDTDTWSEVTRMTSGRSGVGVAVTMEPCRKQIDQQNCTC

Approximately 64 kDa

Greater than 85% as determined by reducing SDS-PAGE

Lyophilized powder.

Lyophilized from a 0.2 μm filtered solution of 20 mM Tris, 500 mM NaCl, 3 mM DTT, 10% Glycerol, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween 80 are added as protectants before lyophilization.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 2 years. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.