Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-DARPP-32 (Ser97) (D11A5) Rabbit mAb #3401

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R Endogenous 32 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
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

Phospho-DARPP-32 (Ser97) (D11A5) Rabbit mAb detects endogenous levels of DARPP-32 only when phosphorylated at Ser97.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser102 of human DARPP-32 protein (equivalent to Ser97 of mouse DARPP-32 protein).

Western Blotting

Western Blotting

Western blot analysis of extracts from MKN-45 cells using Phospho-DARPP-32 (Ser97) (D11A5) Rabbit mAb. The phospho-specificity of the antibody was verified by treating the membrane with (+) or without (-) calf intestinal phosphatase (CIP) after western transfer.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse brain, NIH/3T3 cells, rat brain, and C6 cells using Phospho-DARPP-32 (Ser97) (D11A5) Rabbit mAb (upper) or PKA RI-α (D54D9) Rabbit mAb #5675 (lower) (left panel). The phospho-specificity of the antibody was verified by blocking with a phospho (middle panel) or nonphosphopeptide (right panel).

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).

Drugs of abuse such as cocaine and food reinforcement learning activate the dopamine D1 receptor-signaling cascade. The downstream effector DARPP-32 is dephosphorylated at Ser97 inhibiting its nuclear export. This enables DARPP-32 to function as an inhibitor of protein phosphatase-1, increasing phosphorylation of histone H3 at Ser10. Knock-in mice bearing a DARPP-32 Ser97 to Ala (S97A) mutation demonstrate changed behavioral effects to drugs of abuse and a decreased motivation for food (4).

  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.
  4. Stipanovich, A. et al. (2008) Nature 453, 879-84.

Application References

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

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