Product Pathways - Neuroscience
Phospho-DARPP-32 (Thr75) Antibody #2301
PhosphoSitePlus® protein, site, and accession data: DARPP-32
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | M R | Endogenous | 32 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
M=Mouse
R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 2301:
- Western Blotting
Specificity / Sensitivity
Phospho-DARPP-32 (Thr75) Antibody detects endogenous levels of DARPP-32 only when phosphorylated at threonine 75. The antibody does not cross-react with DARPP-32 phosphorylated at Thr34.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr75 of human DARPP-32. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from rat brain cortex, untreated or phosphorylated in vitro by PKA, MAP kinase or cdc2 kinase, using Phospho-DARPP-32 (Thr75) Antibody #2301 (top), Phospho-DARPP-32 (Thr34) Antibody (middle) or DARPP-32 Antibody #2302 (bottom).
Background
DARPP-32 (dopamine and cyclic AMP-regulated phosphoprotein, relative molecular mass 32,000) is a cytosolic protein highly enriched in medium-sized spiny neurons of the neostriatum (1). It is a bifunctional signaling molecule that controls serine/threonine kinase and serine/threonine phosphatase activity (2). Dopamine stimulates phosphorylation of DARPP-32 through D1 receptors and activation of PKA. PKA phosphorylation of DARPP-32 at Thr34 converts it into an inhibitor of protein phosphatase 1 (1). DARPP-32 is converted into an inhibitor of PKA when phosphorylated at Thr75 by cyclin-dependent kinase 5 (CDK5) (2). Mice containing a targeted deletion of the DARPP-32 gene exhibit an altered biochemical, electrophysiological, and behavioral phenotype (3).
- Nishi, A. et al. (1997) J. Neurosci. 17, 8147-8155.
- Bibb, J.A. et al. (1999) Nature 402, 669-671.
- Fienberg, A.A. et al. (1998) Science 281, 838-842.
Application References
- Danielli, B. et al. (2010) J Neurochem 112, 531-41. Applications: Western Blotting
- Scheggi, S. et al. (2007) J Neurochem 103, 1168-83. Applications: Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 2302 DARPP-32 Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.
