Product Pathways - Neuroscience
Phospho-DARPP-32 (Thr34) Antibody #2304
| Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|
| W | H M R | 32 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-DARPP-32 (Thr34) Antibody detects endogenous levels of DARPP-32 only when phosphorylated at threonine 34. The antibody does not cross-react with DARPP-32 phosphorylated at Thr75. This antibody may also detect PP-1 inhibitor.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr34 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).
IF-F
Confocal microscopic images of mouse retina labeled with DARPP-32 Antibody #2302 (A, red), Phospho-DARPP-32 (Thr34) Antibody #2304 (C) and Phospho-DARPP-32 (Thr75) Antibody #2301 (D) compared to a nonspecific control antibody (B). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
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
- Beaulieu, J.M. et al. (2004) Proc Natl Acad Sci U S A 101, 5099-104. This article references the use of Phospho-DARPP-32 (Thr34) Antibody in the following applications: Western Blotting
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