Product Pathways - Tyrosine Kinase/ Adaptors
Phospho-β-Arrestin 1 (Ser412) (6-24) Mouse mAb #2416
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP | H M R | Endogenous | 50 | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-β-Arrestin 1 (Ser412) (6-24) Mouse mAb detects endogenous levels of β-arrestin 1 only when phosphorylated at serine 412. The antibody does not cross-react with beta-arrestin 2.
Source / Purification
Monoclonal antibody is produced by immunizing mice with a synthetic peptide (KLH-coupled) corresponding to residues surrounding Ser412 of human β-arrestin 1.
Western Blotting
Western blot analysis of extracts from HEK293 cells alone (-), expressing β-arrestin 1 (wt) or β-arrestin (Ser412Ala) mutant (Ala412), using Phospho-β-Arrestin 1 (Ser412) (6-24) Mouse mAb. Lanes 1 and 3 show endogenous levels of phosphorylated β-arrestin 1.
IP
Immunoprecipitation of β-arrestin 1 from HEK293 cells using a polyclonal antibody to phospho-β-arrestin 1 (Ser412), followed by alkaline phosphaphatase (CIP) treatment. β-arrestin 1 was detected by Phospho-beta-Arrestin 1 (Ser412) (6-24) Mouse mAb. CIP treatment abolished the β-arrestin 1 signal, indicating that the monoclonal antibody is phospho-specific.
Background
Arrestin proteins function as negative regulators of G protein coupled receptor (GPCR) signaling. Cognate ligand binding stimulates GPCR phosphorylation, which is followed by binding of arrestin to the phosphorylated GPCR and the eventual internalization of the receptor and desensitization of GPCR signaling (1). Four distinct mammalian arrestin proteins are known. Arrestin 1 (also known as S-arrestin) and arrestin 4 (or X-arrestin) are localized to retinal rods and cones, respectively. Arrestin 2 (also known as β-arrestin 1) and arrestin 3 (or β-arrestin 2) are ubiquitously expressed and bind to most GPCRs (2). β-arrestin proteins function as adapters and scaffold proteins and play important roles in other processes, such as recruiting c-Src family proteins to GPCRs in ERK activation pathways (3,4). β-arrestins are also involved in some receptor tyrosine kinase signaling pathways (5-8). Additional evidence suggests that β-arrestin proteins translocate to the nucleus and help regulate transcription by binding transcriptional cofactors (9,10).
Erk1/2 constitutively phosphorylates β-arrestin 1 at carboxy-terminal Ser412, which promotes cytosolic localization of the scaffold protein (11). Agonist stimulation of β2-adrenergic receptors results in recruitment of β-arrestin 1 to the plasma membrane and rapid dephosphorylation of arrestin. Dephosphorylation is an essential step of β-arrestin 1-mediated receptor endocytosis, but it is not required for receptor desensitization (12).
- Shenoy, S.K. and Lefkowitz, R.J. (2005) Sci STKE 2005, cm10.
- Lefkowitz, R.J. and Shenoy, S.K. (2005) Science 308, 512-7.
- Luttrell, L.M. et al. (1999) Science 283, 655-61.
- Luttrell, L.M. et al. (1999) Curr Opin Cell Biol 11, 177-83.
- Luttrell, L.M. and Lefkowitz, R.J. (2002) J Cell Sci 115, 455-65.
- Waters, C. et al. (2004) Semin Cell Dev Biol 15, 309-23.
- Lefkowitz, R.J. and Whalen, E.J. (2004) Curr Opin Cell Biol 16, 162-8.
- Waters, C.M. et al. (2005) Cell Signal 17, 263-77.
- Kang, J. et al. (2005) Cell 123, 833-47.
- Ma, L. and Pei, G. (2007) J Cell Sci 120, 213-8.
- Lin, F.T. et al. (1999) J Biol Chem 274, 15971-4.
- Lin, F.T. et al. (1997) J Biol Chem 272, 31051-7.
Application References
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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.