Application

Research Sub CategoryNuclear Receptors

Western Blotting Analysis: A representative lot detected endogenous MafK as well as Zn+2-induced expression of exogenous MafK using nuclear extracts from a mouse erythroleukemia (MEL) cell line transfected with MafK cDNA in a metallothionein IIA gene promoter-driven expression plasmid (Igarashi, K., et al. (1995). Proc Natl Acad Sci U S A. 92(16):7445-7449).
Chromatin Immunoprecipitation Analysis: A representative lot detected MafK association with the GST-P enhancer (GPE1) in rat hepatoma cells (H4IIE and dRLh84) and hyperplastic nodules/HN-bearing pre-neoplastic rat hepatocytes, but not in normal rat hepatocytes. While MafK association with the NAD(P)H:quinone oxidoreductase 1 (NQO1) antioxidant responsive element (ARE) was detected in all four sample types (Ikeda, H., et al. (2004). Biochem J. 380(Pt 2):515-521).
Electrophoretic Mobility Shift Assay Analysis: A representative lot caused a supershift of MafK-containing complex in EMSA using radiolabeled oligonucleotides containing PBGD gene NF-E2 binding site and nuclear extracts from a mouse erythroleukemia (MEL) cell line induced to overexpress MafK (Igarashi, K., et al. (1995). Proc Natl Acad Sci U S A. 92(16):7445-7449).
Electrophoretic Mobility Shift Assay (EMSA): A representative lot caused a supershift of both the constitutive and tBHQ-induced ARE-binding complex (Complex I and II, respectively) in EMSA using a radiolabeled oligonucleotide containing antioxidant responsive element (ARE) NF-E2 binding site and nuclear extracts from untreated and tBHQ-treated K562 cells (Kim, Y.C., et al. (2003). Oncogene. 22(12):1860-1865).

Research CategoryEpigenetics & Nuclear Function

This Anti-MafK/Nfe2u Antibody is validated for use in Western Blotting, Chromatin Immunoprecipitation (ChIP) and Electrophoretic Mobility Shift Assay for the detection of MafK/Nfe2u.

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

Transcription factor MafK (UniProt Q61827; also known as Erythroid transcription factor NF-E2 p18 subunit, Nuclear factor erythroid derived 2 ubiquitous, p18 NF-E2) is encoded by the Mafk (also known as Nfe2u, AW061068) gene (Gene ID 17135) in murine species. MafK, MafG and MafF consistute a family of transcription factors that dimerize with the CNC (cap′ n′ collar) family regulatory protein Nrf2 (NF-E2 p45-related factor 2) via their bZip domain. The Nrf2/small Maf heterodimer regulates downstream genes, including those of phase II detoxifying enzymes, by targeting antioxidant responsive element (ARE) and Maf recognition element (MARE).

Immunogen

Maltose binding protein-tagged recombinant protein corresponding to mouse MafK/Nfe2u.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Rabbit polyclonal serum with 0.05% sodium azide.

Unpurified

Quality

Evaluated by Western Blotting in MEL (mouse erythroleukemia) cell lysate.

Western Blotting Analysis: 1:2,000 dilution of this antibody detected MafK/NF-E2 in 20 µg of MEL (mouse erythroleukemia) cell lysate.

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

~18 kDa observed. Uncharacterized band(s) may appear in some lysates.