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Flag nrf2

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Flag-NRF2 is a recombinant protein that contains the Flag epitope tag fused to the Nrf2 (Nuclear Factor Erythroid 2-Related Factor 2) protein. The Flag tag allows for easy detection and purification of the Nrf2 protein.

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Lab products found in correlation

Flag keap1, by Addgene (3 mentions) Profection, by Promega (2 mentions) Viafect, by Promega (2 mentions) Ha k48, by Addgene (2 mentions) Ha k63, by Addgene (2 mentions) Gfp polyq74, by Addgene (2 mentions) Gfp p62, by Addgene (2 mentions) Gfp nrf2, by Addgene (2 mentions) Gfp keap1, by Addgene (2 mentions) Myc nrf2, by Addgene (2 mentions) Flag ulk1, by Addgene (2 mentions) His k63, by Thermo Fisher Scientific (2 mentions) Effectene, by Qiagen (2 mentions) Pcdna3 egfp c4 nrf2, by Addgene (1 mentions) Pgl4.74 renilla luciferase, by Promega (1 mentions) Pcdna3.1 flag nrf2, by Addgene (1 mentions)

Close spelling variants of this product

PCDNA3.1 FLAG NRF2 NC16 pCDNA3.1 FLAG NRF2

3 protocols using flag nrf2

1

Molecular Mechanisms of NRF2 Regulation

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GFP‐polyQ74 (#40262), GFP‐p62 (#38277), GFP‐NRF2 (#21549), GFP‐KEAP1 (#28025), Flag‐KEAP1 (#28023), Myc‐NRF2 (#21555), Flag‐NRF2 (#36971), HA‐K48 (#17605), HA‐K63 (#17606), Flag‐ULK1 (#27636) were procured from Addgene. GFP‐TRIM16 and Flag‐TRIM16 were cloned as described previously (Chauhan et al, 2016). Myc‐TRIM16, Myc‐TRIM16 deletion constructs, and His‐K63‐UB were generated using Gateway cloning strategy as per standard protocol (Invitrogen).
The siRNA for NRF2 and p62 were purchased from Sigma, and TRIM16, Ubb, Ube2n siRNA were from Dharmacon. For overexpression experiments, HEK293T cells were transfected using calcium phosphate method as per the manufacturer's instructions (Profection, Promega). Other cells are transfected using Effectene (Qiagen) or Viafect (Promega) or Interference (Polyplus) as per the manufacturer's instruction.
Jena K.K., Kolapalli S.P., Mehto S., Nath P., Das B., Sahoo P.K., Ahad A., Syed G.H., Raghav S.K., Senapati S., Chauhan S, & Chauhan S. (2018). TRIM16 controls assembly and degradation of protein aggregates by modulating the p62‐NRF2 axis and autophagy. The EMBO Journal, 37(18), e98358.
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2

Plasmids for NRF2 and HIF1α Regulation

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For transient transfections the following plasmids were used: pGL3-8xARE42 (link), or pGLHIF1.343 (link), pGL4.74 renilla luciferase (#E6921, Promega), pGFP-NRF2 (pcDNA3-EGFP-C4-Nrf2, #21549, Addgene), Flag-NRF2 (pCDNA3.1 FLAG NRF2, #36971, Addgene), Flag-Keap1 (#28023, Addgene), p(HA)HIF1alpha (P402A,P564A) (#52636, Addgene). To generate plasmid GFP-NRF2-S40A, a modified QuickChange protocol was used44 (link) to perform a PCR using pGFP-NRF2 as the template and primers NRF2-S40A-for (5ʹ-GTCGAGAAGTATTTGACTTCGCCCAGCGACGGAAA-GAGTATGAGCTGGAAAA-3ʹ) and NRF2-S40A-rev (5ʹ-GGCGAAGTCAAATAC-TTCTCGACTTACTCCAAGA TCTATATCTTGCCTCCAAAGTATGTCA-3ʹ). pLKO.1-shRNA-NRF2 tet-on (tetracycline inducible) contained the sequence 5ʹ-GCTCCTACTGTGATGTGAAAT-3ʹ of NRF2 mRNA (GenBank acc. no. NM_001145412), pLKO.1-shRNA-PERK tet-on (tetracycline inducible) contained the sequence 5ʹ-GGAACGACCTGAAGCTATAAA-3ʹ of PERK mRNA (GenBank acc. no. NM_004836).
Küper A., Baumann J., Göpelt K., Baumann M., Sänger C., Metzen E., Kranz P, & Brockmeier U. (2021). Overcoming hypoxia-induced resistance of pancreatic and lung tumor cells by disrupting the PERK-NRF2-HIF-axis. Cell Death & Disease, 12(1), 82.
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3

Characterization of NRF2-KEAP1 Pathway

Cited 1 time
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GFP-polyQ74 (#40262), GFP-p62 (#38277), GFP-NRF2 (#21549), GFP-KEAP1 (#28025), Flag-KEAP1 (#28023), Myc-NRF2 (#21555), Flag-NRF2 (#36971), HA-K48 (#17605), HA-K63 (#17606), Flag-ULK1 (#27636) were procured from Addgene. GFP-TRIM16 and Flag-TRIM16 were cloned as described previously (Chauhan et al, 2016 (link)). Myc-TRIM16, Myc-TRIM16 deletion constructs, and His-K63-UB were generated using Gateway cloning strategy as per standard protocol (Invitrogen).
The siRNA for NRF2 and p62 were purchased from Sigma, and TRIM16, Ubb, Ube2n siRNA were from Dharmacon. For overexpression experiments, HEK293T cells were transfected using calcium phosphate method as per the manufacturer’s instructions (Profection, Promega). Other cells are transfected using Effectene (Qiagen) or Viafect (Promega) or Interference (Polyplus) as per the manufacturer’s instruction.
Jena K.K., Kolapalli S.P., Mehto S., Nath P., Das B., Sahoo P.K., Ahad A., Syed G.H., Raghav S.K., Senapati S., Chauhan S, & Chauhan S. (2018). TRIM16 controls assembly and degradation of protein aggregates by modulating the p62‐NRF2 axis and autophagy. The EMBO Journal, 37(18), e98358.
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