Cell Signaling Technology

Product Pathways - Neuroscience

DARPP-32 Antibody #2302

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. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

DARPP-32 Antibody detects endogenous levels of total DARPP-32 independent of phosphorylation. This antibody may also detect PP1.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr34 of human DARPP-32. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

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

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).

  1. Nishi, A. et al. (1997) J. Neurosci. 17, 8147-8155.
  2. Bibb, J.A. et al. (1999) Nature 402, 669-671.
  3. Fienberg, A.A. et al. (1998) Science 281, 838-842.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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