Application

Research Sub CategoryTranscription Factors

Electrophoretic Mobility Shift Assay (EMSA): A representative lot caused a supershift of DNA-NRF-1 complex in EMSA using radiolabeled oligonucleotide containing human mitochondrial transcription specificity factor genes (TFB1M & TFB2M) NRF-1-binding site and HeLa nuclear extract (NE) or purified recombinant NRF-1 (Gleyzer, N., et al. (2005). Mol. Cell. Biol. 25(4):1354-1366).
Electrophoretic Mobility Shift Assay (EMSA): A representative lot caused a supershift of DNA-NRF-1 complex in EMSA using radiolabeled oligonucleotide containing rat cytochrome C NRF-1-binding site and HeLa nuclear extract (NE), purified HeLa NRF-1 or recombinant NRF-1. No shift by this antiserum of HeLa NE proteins complexed with oligonucleotide containing mouse COXVb gene NRF-2-binding site was observed (Virbasius, C.A., et al. (1993). Genes Dev. 7(12A):2431-2445).

Research CategoryEpigenetics & Nuclear Function

Anti-NRF-1 Antibody is an antibody against NRF-1 for use in Western Blotting, Electrophoretic Mobility Shift Assay.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Nuclear respiratory factor 1 (UniProt Q16656; also known as Alpha-pal, Alpha palindromic-binding protein, NRF-1) is encoded by the NRF1 (Also known as ALPHA-PAL) gene (Gene ID 4899) in human. NRF-1 is a redox-responsive transcription factor that plays an anti-oxidant, pro-survival role in mitochondrial biogenesis process. NRF-1 is upregulated and activated/phosphorylated by Akt in response to oxidant exposure or increased cellular reactive oxygen species (ROS), reulsting in NRF-1 nuclear translocation, where it binds to DNA as a homodimer and mediates the transcription of mitochondrial transcription factor A (mtTFA) that is needed for mitochondrial DNA transcription.

Immunogen

Recombinant human NRF-1.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Goat polyclonal serum with 0.05% sodium azide.

Unpurified

Quality

Evaluated by Western Blotting in HeLa nuclear extract.

Western Blotting Analysis: An 1:200 dilution of this antibody detected NRF1 in 10 µg of HeLa nuclear extract.

Specificity

Expected to react with all four spliced isoforms of human NRF-1 reported by UniProt (Q16656). This antiserum bound DNA-NRF-1 complex as evidenced by a specific supershift of NRF-1, but not NRF-2, DNA complex in EMSA (Virbasius, C.A., et al. (1993). Genes Dev. 7(12A):2431-2445).

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~68 kDa observed. 53.54 kDa (isoform Long), 46.16 kDa (isoform Short), 36.05 kDa (isoform 3), 55.67 kDa (isoform 4) calculated.