Cell Signaling Technology

Product Pathways - Nuclear Receptor Signaling

Phospho-Rev-erbα (Ser55/59) Antibody #2129

Applications Reactivity Sensitivity MW (kDa) Source
W H (M) (R) (B) Transfected Only 80 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  B=Bovine
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Rev-erbα (Ser55/59) Antibody detects transfected levels of Rev-erbα protein when phosphorylated at Ser55/59. The antibody does not cross-react with other nuclear receptor proteins, including Rev-erbβ.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser55 and Ser59 of human Rev-erbα. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, either untransfected or transfected with DYKDDDDK-tagged Rev-erbα expression constructs, using Phospho-Rev-erbα (Ser55/Ser59) Antibody (upper) or DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368 (lower). The phospho-specificity of the antibody was verified by transfecting cells with a mutant DYKDDDDK-Rev-erbα (Ser55/59Ala) expression construct and by incubating 293/Rev-erbα (WT) extracts with (+) or without (-) λ phosphatase.

Background

Reverse orientation c-erbA gene α (Rev-erbα, EAR-1, or NR1D1) is a widely expressed member of the orphan nuclear receptor family of proteins (1). Rev-erbα is highly expressed in adipose tissue, skeletal muscle, brain and liver, and regulates cellular proliferation and differentiation. Expression increases during differentiation in adipocytes and ectopic expression of Rev-erbα potentiates the adipocyte differentiation of 3T3-L1 cells (2). In addition, expression oscillates with circadian rhythm in liver cells and Rev-erbα regulates expression of BMAL1, ApoA-I and ApoC-III, all key regulators of circadian rhythm (3,4,5,6,7). Phosphorylation of Rev-erbα Ser55 and Ser59 by GSK3β appears to stabilize Rev-erbα protein levels and is important for synchronizing and maintaining the circadian clock (8). Rev-erbα also regulates inflammation by targeting the NF-κB responsive genes IL-6 and COX-2 (9). Rev-erbα lacks the activation function 2 domain required for ligand-dependent activation of transcription by other members of the nuclear receptor family; thus it behaves as a constitutive repressor protein, recruiting the nuclear receptor co-repressor (N-CoR)/HDAC3 complex to target genes to repress transcription (10).

  1. Harding, H.P. and Lazar, M.A. (1993) Mol. Cell Biol. 13, 3113-3121.
  2. Chawla, A. and Lazar, M.A. (1993) J. Biol. Chem. 268, 16265-16269.
  3. Torra, I.P. et al. (2000) Endocrinology 141, 3799-3806.
  4. Preitner, N. et al. (2002) Cell 110, 251-260.
  5. Vu-Dac, N. et al. (1998) J. Biol .Chem. 273, 25713-25720.
  6. Coste, H. and Rodríguez, J.C. (2002) J. Biol. Chem. 277, 27120-27129.
  7. Raspé, E. et al. (2002) J. Lipid Res. 43, 2172-2179.
  8. Yin, L. et al. (2006) Science 311, 1002-1005.
  9. Migita, H. et al. (2004) FEBS Lett. 561, 69-74.
  10. Yin, L. and Lazar, M.A. (2005) Mol. Endocrinol. 19, 1452-1459.

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