Product Pathways - Neuroscience
Phospho-DARPP-32 (Thr75) Antibody #2301
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IF-F | H M R | Endogenous | 32 | Rabbit |
Applications Key:
W=Western Blotting
IF-F=Immunofluorescence (Frozen)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Protocols
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 rabbits with a synthetic phospho-peptide (KLH-coupled) 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).
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
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Companion Products
- 2302 DARPP-32 Antibody
- 2304 Phospho-DARPP-32 (Thr34) 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
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.
