PRODUCT CODE: ET1608-23

DARPP32 Recombinant Rabbit Monoclonal Antibody [SU0329] (ET1608-23)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of DARPP32 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-23, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Mouse brain tissue lysate<br />
Lane 2: Rat brain tissue lysate
  • Western blot analysis of DARPP32 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-23, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Mouse brain tissue lysate<br />
Lane 2: Rat brain tissue lysate
  • ICC staining of DARPP32 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of DARPP32 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-DARPP32 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-DARPP32 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-DARPP32 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of DARPP32 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-23, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of DARPP32 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-23, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Mouse brain tissue lysate
Lane 2: Rat brain tissue lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

DARPP32 Recombinant Rabbit Monoclonal Antibody [SU0329] (ET1608-23)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

SH-SY-5Y, MCF-7, mouse brain tissue, rat brain tissue, human gastric carcinoma tissue, human breast carcinoma tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SU0329

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

32 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

DARPP32

SYNONYMS

DARPP 32 antibody; DARPP-32 antibody; Dopamine and cAMP regulated neuronal phosphoprotein 32 antibody; Dopamine and cAMP regulated neuronal phosphoprotein antibody; Dopamine and cAMP regulated phosphoprotein antibody; Dopamine and cAMP regulated phosphoprotein DARPP 32 antibody; Dopamine and cAMP regulated phosphoprotein DARPP32 antibody; Dopamine- and cAMP-regulated neuronal phosphoprotein antibody; FLJ20940 antibody; IPPD antibody; Neuronal phosphoprotein DARPP 32 antibody; PPP1R1B antibody; PPR1B_HUMAN antibody; Protein phosphatase 1 regulatory (inhibitor) subunit 1B antibody; Protein phosphatase 1 regulatory inhibitor subunit 1B antibody; Protein phosphatase 1 regulatory subunit 1B antibody

SEQUENCE SIMILARITIES

Belongs to the protein phosphatase inhibitor 1 family.

POST-TRANSLATIONAL MODIFICATION

Dopamine- and cyclic AMP-regulated neuronal phosphoprotein.; Phosphorylation of Thr-34 is required for activity.

SUBCELLULAR LOCATION

Cytoplasm.

FUNCTION

Dopaminergic signaling pathways, which are essential for multiple brain functions, are abnormal in several neurological disorders, such as schizophrenia, Parkinson's disease and drug abuse. DARPP-32 (for dopamine and adenosine 3',5'-monophosphate-regulated phosphoprotein) is abundant in neurons that receive dopaminergic input. Activation of PKA and the consequent phosphorylation of DARPP-32 on threonine occurs in response to dopamine acting upon D1-like receptors. Dopamine interaction with D2-like receptors results in the inhibition of PKA activation, the activation of protein phosphatase 2B and the consequent dephosphorylation of DARPP-32. Neurotransmitters other than dopamine may also be able to stimulate the phosphorylation or dephosphorylation of DARPP-32. Phosphorylated DARPP-32 is a potent inhibitor of PP-1.