WB result of DARPP32 Recombinant Rabbit mAb
Primary antibody: DARPP32 Recombinant Rabbit mAb at 1/4000 dilution
Lane 1: mouse brain lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 23 kDa
Observed MW: 32 kDa
DARPP32 Recombinant Rabbit mAb (S-1637-62)
DARPP32 Recombinant Rabbit mAb (S-1637-62)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | DARPP32 |
| Synonyms | Protein phosphatase 1 regulatory subunit 1B; DARPP-32; Dopamine- and cAMP-regulated neuronal phosphoprotein; PPP1R1B |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm |
| Accession | Q9UD71 |
| Clone Number | S-1637-62 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, IF |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | Human brain, mouse brain, rat brain |
| Predicted Reactivity | Bv, Pg |
| Purification | Protein A |
| Concentration | 2 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:4000 | Ms |
| IHC-P | 1:100-1:250 | Hu, Ms, Rt |
| IF | 1:400 | Ms |
Background
DARPP32, or dopamine- and cAMP-regulated phosphoprotein with an apparent molecular weight of 32 kDa, is a signaling protein that plays a crucial role in the brain, particularly in regions rich in dopaminergic nerve terminals. It is known to be a potent inhibitor of protein phosphatase 1 (PP1) when phosphorylated at Thr34 by cAMP-dependent protein kinase (PKA). DARPP32 is also phosphorylated at other sites by various kinases, which modulates its properties. For instance, phosphorylation at Thr75 by Cdk5 converts DARPP32 into an inhibitor of PKA. This dual functionality makes DARPP32 a key integrator in signal transduction pathways. DARPP-32 is involved in the regulation of a range of physiological and behavioral responses to various stimuli, including neurotransmitters like dopamine and glutamate, as well as drugs of abuse such as cocaine, amphetamine, nicotine, and caffeine. The phosphorylation state of DARPP-32 is intricately regulated, with phosphorylation at Ser102 and Ser137 by CK2 and CK1, respectively, influencing its activity. This complex regulation enables DARPP-32 to modulate the activity of PP-1 and PKA, which in turn affects electrophysiological, transcriptional, and behavioral responses. Moreover, DARPP-32 has been implicated in several psychiatric and neurological disorders, including schizophrenia, and its expression has been observed in a variety of epithelial cells beyond the central nervous system. High levels of DARPP-32 have been associated with survival in certain cancers, suggesting it may play a role in tumorigenesis and could be a potential therapeutic target.
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Western Blot
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-DARPP32 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-DARPP32 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung cancer. Anti-DARPP32 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse cerebral cortex. Anti-DARPP32 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat cerebral cortex. Anti-DARPP32 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunofluorescence
IF shows positive staining in paraffin-embedded mouse cerebral cortex. Anti- DARPP32 antibody was used at 1/400 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
