Phospho-Nur77 (Ser351) (D22G5)

5095:

Western Blotting

Phospho-Nur77 (Ser351) (D22G5) Rabbit mAb detects endogenous levels of Nur77 protein only when phosphorylated at Ser351.

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser351 of human Nur77 protein.

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with TPA #4174 and calcium ionophore A23187 for the indicated times, using Phospho-Nur77 (Ser351) (D22G5) Rabbit mAb (upper) or Nur77 (D63C5) XP^® Rabbit mAb #3960 (lower).

Western blot analysis of extracts from Jurkat cells, untreated or treated with TPA #4174 and calcium ionophore A23187 for 4 hours, using Phospho-Nur77 (Ser351) (D22G5) Rabbit mAb. Antibody phospho-specificity was confirmed by pre-incubating the antibody with either a phosphorylated site-specific peptide to block the signal or a non-phosphorylated site-specific peptide that does not block the signal.

Nur77, also known as TR3 and NGFI-B, is an immediate-early response gene and an orphan member of the steroid/thyroid/retinoid receptor superfamily (1-3). Nur77 is composed of an amino-terminal transactivation domain, a central DNA-binding domain and a carboxy-terminal ligand-binding domain. Expression of Nur77 is rapidly induced by a variety of stimuli, including apoptotic, mitogenic and stress signals (1-6). It has been proposed to have many functions related to cell proliferation, differentiation and apoptosis. Nur77 has been extensively studied in T cells where it has been implicated in the process of negative selection and TCR-mediated apoptosis (5,6). Nur77 binds to specific DNA elements leading to the regulation of target genes (7). As a possible mechanism for regulating apoptosis, Nur77 can induce the expression of apoptotic genes such as FasL and TRAIL (8,9). Nur77 is heavily phosphorylated by multiple kinases, which may affect its transactivation activity as well as its subcellular localization (4,10,11). Translocation of Nur77 from the nucleus to the mitochondria can regulate its association with Bcl-2 and control the release of cytochrome c, thereby triggering apoptosis (12,13).

Phosphorylation of Nur77 by Akt or RSK occurs at Ser351 (corresponding to rat Nur77 Ser350 and Ser354 of mouse Nur77), a site within the Nur77 DNA binding domain (14-16). Serine phosphorylation at this site can down regulate transcriptional activity of Nur77 (10,17).

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2. Chang, C. and Kokontis, J. (1988) Biochem. Biophys. Res. Commun. 155, 971-977.
3. Milbrandt, J. (1988) Neuron 1, 183-188.
4. Fahrner, T.J. et al. (1990) Mol. Cell. Biol. 10, 6454-6459.
5. Liu, Z.G. et al. (1994) Nature 367, 281-284.
6. Woronicz, J.D. et al. (1994) Nature 367, 277-281.
7. Wilson, T.E. et al. (1991) Science 252, 1296-1300.
8. Weih, F. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 5533-5538.
9. Rajpal, A. et al. (2003) EMBO J. 22, 6526-6536.
10. Hirata, Y. et al. (1993) J. Biol. Chem. 268, 24808-24812.
11. Hazel, T.G. et al. (1991) Mol. Cell. Biol. 11, 3239-3246.
12. Li, H. et al. (2000) Science 289, 1159-1164.
13. Lin, B. et al. (2004) Cell 116, 527-540.
14. Pekarsky, Y. et al. (2001) Proc. Natl. Acad. Sci. U S A 98, 3690-3694.
15. Han, Y.H. et al. (2006) Oncogene 25, 2974-2986.
16. Wingate, A.D. et al. (2006) Biochem. J. 393, 715-724.
17. Katagiri, Y. et al. (1997) J. Biol. Chem. 272, 31278-31284.

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• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7727 Biotinylated Protein Ladder Detection Pack
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 3960 Nur77 (D63C5) XP^® Rabbit mAb

For Research Use Only. Not For Use In Diagnostic Procedures.