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Transam nrf2 assay

Manufactured by Active Motif
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The TransAm Nrf2 assay is a sensitive and specific kit for the detection and quantification of activated Nrf2 in cell or tissue samples. The assay utilizes a 96-well plate format and provides a simple, non-radioactive method for measuring Nrf2 DNA-binding activity.

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Lab products found in correlation

Elx800 microplate reader, by Agilent Technologies (1 mentions) Infinite m200 pro plate reader, by Tecan (1 mentions)

Close spelling variants of this product

TransAM Nrf2 Transcription Factor Assay Kit TransAM NRF2 assay Kit TransAM Nrf2

3 protocols using transam nrf2 assay

1

Quantification of Nrf2 Transcriptional Activity

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Nrf2 activity was determined with the TransAm Nrf2 assay (Active Motif, Carlsbad, CA). Nuclear extracts (10 μg) were incubated with immobilized oligonucleotide containing the ARE consensus binding site (5´-GTCACAGTGACTCAGCAGAATCTG-3′) on 96-well plates. The active form of Nrf2 that bound the oligo was detected using primary anti-Nrf2 antibody after treating with secondary HRP conjugated antibody. As a result of the specific transcription factor activity in nuclear extracts, the chromogen formed was determined using a Synergy multi-modem plate reader (BioTek, Winooski, VT) at 450 nm, and expressed as Nrf2/ARE binding activity.
Colonnello A., Aguilera-Portillo G., Rubio-López L.C., Robles-Bañuelos B., Rangel-López E., Cortez-Núñez S., Evaristo-Priego Y., Silva-Palacios A., Galván-Arzate S., García-Contreras R., Túnez I., Chen P., Ashner M, & Santamaría A. (2019). Comparing the Neuroprotective Effects of Caffeic Acid in Rat Cortical Slices and Caenorhabditis elegans: Involvement of Nrf2 and SKN-1 Signaling Pathways. Neurotoxicity research, 37(2), 326-337.
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2

Nrf2 Transcriptional Activity Assay

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Nrf2 activity was determined with the TransAM Nrf2 assay (Active Motif, Carlsbad, CA) according to the manufacturer's instructions [44 (link)]. Briefly, nuclear extracts from different groups were incubated with ARE consensus site oligonucleotides (5′-GTCACAGTGACTCAGCAGAATCTG-3′) immobilized to 96-well plates. Bound protein was detected with an antibody specific to Nrf2 and visualized by colorimetric reaction catalyzed by HRP-conjugated secondary antibody. The absorbance was measured at 405 nm using an ELX800 microplate reader (Bio-Tek Instruments, Winooski, VT, USA).
Sun X., Xie Z., Hu B., Zhang B., Ma Y., Pan X., Huang H., Wang J., Zhao X., Jie Z., Shi P, & Chen Z. (2019). The Nrf2 activator RTA-408 attenuates osteoclastogenesis by inhibiting STING dependent NF-κb signaling. Redox Biology, 28, 101309.
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3

Quantifying NRF2 Transcriptional Activity

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NRF2 activity was determined using the TransAM NRF2 assay (50296, Active Motif) according to the manufacture’s protocol. Briefly, nuclear extracts (5 μg) were incubated with ARE consensus site oligonucleotides (5′-GTCACAGTGACTCAGCAGAATCTG-3′); immobilized to 96-well plates. Bound protein was detected with an antibody specific to DNA-bound NRF2 and visualized by colorimetric reaction catalyzed by horseradish peroxidase-conjugated secondary antibody, and absorbance was measured at 405 nm using the Infinite M200 Pro plate reader (Tecan).
Shashank S., Arpita C., Kosmacek E.A., Davis P.J., Tyson M.J, & Oberley-Deegan R.E. (2020). Manganese porphyrin, MnTE-2-PyP, treatment protects the prostate from radiation-induced fibrosis (RIF) by activating the NRF2 signaling pathway and enhancing SOD2 and sirtuin activity. Free radical biology & medicine, 152, 255-270.
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