Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Mouse IgG1

UniProt ID:

#P49407

Entrez-Gene Id:

408

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser412 of human β-arrestin 1.

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 (X-arrestin) are localized to retinal rods and cones, respectively. Arrestin 2 (also known as β-arrestin 1) and arrestin 3 (β-arrestin 2) are ubiquitously expressed and bind to most GPCRs (2). β-arrestins function as adaptor 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-arrestins 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).

  1. Shenoy, S.K. and Lefkowitz, R.J. (2005) Sci STKE 2005, cm10.
  2. Lefkowitz, R.J. and Shenoy, S.K. (2005) Science 308, 512-7.
  3. Luttrell, L.M. et al. (1999) Science 283, 655-61.
  4. Luttrell, L.M. et al. (1999) Curr Opin Cell Biol 11, 177-83.
  5. Luttrell, L.M. and Lefkowitz, R.J. (2002) J Cell Sci 115, 455-65.
  6. Waters, C. et al. (2004) Semin Cell Dev Biol 15, 309-23.
  7. Lefkowitz, R.J. and Whalen, E.J. (2004) Curr Opin Cell Biol 16, 162-8.
  8. Waters, C.M. et al. (2005) Cell Signal 17, 263-77.
  9. Kang, J. et al. (2005) Cell 123, 833-47.
  10. Ma, L. and Pei, G. (2007) J Cell Sci 120, 213-8.
  11. Lin, F.T. et al. (1999) J Biol Chem 274, 15971-4.
  12. Lin, F.T. et al. (1997) J Biol Chem 272, 31051-7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#2416

Phospho-β-Arrestin 1 (Ser412) (6-24) Mouse mAb

Western Blotting Image 1: Phospho-β-Arrestin 1 (Ser412) (6-24) Mouse mAb Expand Image
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.
Immunoprecipitation Image 1: Phospho-β-Arrestin 1 (Ser412) (6-24) Mouse mAb Expand Image
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.