Anti-DARPP32 Rabbit Monoclonal Antibody for WB, ELISA - Q9UD71

Core Product Specifications and Parameters

Biological Background: DARPP32 (PPP1R1B) Function and Localization

• DARPP32, encoded by the PPP1R1B gene, is a bifunctional signal transduction molecule that integrates dopamine and cAMP signals through its regulation of protein phosphatase 1 (PP1) and protein kinase A (PKA).
• It acts as a potent inhibitor of PP1 when phosphorylated at Thr34 by PKA, leading to altered phosphorylation of downstream neuronal substrates and modulation of synaptic plasticity.
• The protein is also phosphorylated at Thr75 by cyclin-dependent kinase 5 (CDK5), which converts it into a PKA inhibitor, demonstrating its role as a molecular switch in striatal neurons.
• DARPP32 is predominantly cytoplasmic and highly enriched in medium spiny neurons of the striatum, where it mediates the effects of dopamine, glutamate, and other neurotransmitters.
• Alternative splicing of the PPP1R1B transcript yields multiple isoforms, including a well-characterized full-length form and a truncated variant, which may have distinct functional properties.
• Post-translational modifications such as acetylation and phosphorylation at multiple sites fine-tune its activity, stability, and subcellular distribution.
• Dysregulation of DARPP32 signaling has been associated with neurological and psychiatric disorders, including Parkinson’s disease, schizophrenia, and drug addiction.
• The theoretical molecular weight of the canonical isoform is approximately 23 kDa, consistent with the observed band in Western blotting.

Experimental Guidance and Technical Tips

• For Western blot, use freshly prepared RIPA buffer containing phosphatase and protease inhibitors to preserve the phosphorylation status of DARPP32.
• Consider using a reducing loading buffer and heating samples at 70°C for 10 minutes to avoid aggregation artifacts commonly observed with DARPP32.
• The immunogen corresponds to the N-terminal region (aa1-100); this antibody should recognize both full-length and certain N-terminal fragments.
• Due to high sequence conservation, validate antibody specificity in knockout or siRNA knockdown samples, especially when working with mouse or rat tissues.
• For ELISA, determine optimal coating concentration via checkerboard titration using the immunizing peptide or recombinant DARPP32 protein as standard.
• Include a known positive control (e.g., striatal tissue lysate) and a negative control (e.g., non-neuronal cell line) in each experiment.
• Because DARPP32 is a low-abundance phosphoprotein in some samples, signal enhancement strategies like using high-sensitivity ECL substrates may be necessary for detection.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your comprehensive partner for IVD raw material sourcing, providing validated antibodies, antigen-grade proteins, and ancillary reagents essential for assay development. This anti-DARPP32 rabbit monoclonal antibody supports neuroscience research and diagnostic applications targeting dopamine/cAMP signaling pathways. Leverage our supply capabilities for bulk ordering, customized formulations, and rare target sourcing to accelerate your project from concept to clinic.