Cell Signaling Technology

Product Pathways - Nuclear Receptor Signaling

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

EAR1   HREV   NR1D1   Rev-erbA-alpha   THRAL   V-erbA related protein EAR-1  

No. Size Price
2129S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
2129 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Transfected Only 80 Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat, Bovine,

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.当丝氨酸(55/59位)磷酸化时,Phospho-Rev-erbα (Ser55/59) Antibody能够检测转染水平的Rev-erbα蛋白。该抗体不与其他核受体蛋白包括Rev-erbβ,发生交叉反应。

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.该多克隆抗体通过用合成磷酸肽免疫动物制备,该合成磷酸肽是人Rev-erbα丝氨酸(55和59位)附近的残基。抗体是由蛋白A和肽亲和层析纯化。

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.Western blot方法检测293细胞提取物,细胞不转染或用DYKDDDDK-标记的表达Rev-erbα的构建体转染,使用的抗体为Phospho-Rev-erbα (Ser55/Ser59) Antibody (上图)或DYKDDDDK Tag Antibody (同Sigma's Anti-FLAG® M2 Antibody一样,结合同一抗原表位) #2368 (下图). 抗体的磷酸化特异性由转染的细胞进行验证,细胞用突变的DYKDDDDK-Rev-erbα (Ser55/59Ala)表达构建体转染,293/Rev-erbα (WT )细胞提取物在含(+)或不含(-)λ磷酸酶中孵育。

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).反转定向c-erbA基因α(Rev-erbα, EAR-1或 NR1D1)是一种广泛表达的孤儿核受体蛋白家族成员(1)。Rev-erbα在脂肪组织、骨骼肌、脑和肝中高表达,并调节细胞的增殖和分化。在脂肪细胞分化过程中表达增加,REV-erbα异位表达会增强3T3-L1细胞向脂肪细胞分化(2)。此外,REV-erbα在肝细胞内按昼夜节律振荡表达,并调控BMAL1、ApoA-I和ApoC-III,所有关键调节器都是昼夜节律表达(3,4,5,6,7)。由GSK3β磷酸化REV-erbα的丝氨酸(55和59位),表现出稳定的REV-erbα蛋白质水平,在生物钟的同步和维持中很重要(8)。同样,REV-erbα针对NF-κB应答基因IL-6和COX-2调节炎症(9)。 REV-erbα缺乏激活功能2结构域,这对核受体其他家族成员的配体依赖型转录激活是需要的;因此它作为阻遏蛋白起作用,招募核受体辅阻遏物(N-COR)/ HDAC3复合物的靶基因,抑制转录(10)。

  1. Harding, H.P. and Lazar, M.A. (1993) Mol Cell Biol 13, 3113-21.
  2. Chawla, A. and Lazar, M.A. (1993) J Biol Chem 268, 16265-9.
  3. Torra, I.P. et al. (2000) Endocrinology 141, 3799-806.
  4. Preitner, N. et al. (2002) Cell 110, 251-60.
  5. Vu-Dac, N. et al. (1998) J Biol Chem 273, 25713-20.
  6. Coste, H. and Rodríguez, J.C. (2002) J Biol Chem 277, 27120-9.
  7. Raspé, E. et al. (2002) J Lipid Res 43, 2172-9.
  8. Yin, L. et al. (2006) Science 311, 1002-5.
  9. Migita, H. et al. (2004) FEBS Lett 561, 69-74.
  10. Yin, L. and Lazar, M.A. (2005) Mol Endocrinol 19, 1452-9.

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