DARPP32 Recombinant Rabbit Monoclonal Antibody [SU0329]
cat.: ET1608-23
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IP, FC, IHC-P, IHC-Fr
Clonality: Monoclonal
Clone number: SU0329
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 23 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human DARPP32 aa 1-94 / 204.
Positive control: Mouse striatum tissue, mouse forebrain tissue, human brain tissue, human colon tissue, mouse brian tissue, mouse epididymis tissue, rat brian tissue, Human brain tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Rat hippocampus tissue lysate, SH-SY5Y, MCF-7, Hela.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IHC-P
  IHC-Fr
1:1,000-1:5,000
1:50-1:200
1:50-1:100
1:500-1:1,000
1:500
Uniprot #: SwissProt: Q9UD71 Human | Q60829 Mouse | Q6J4I0 Rat
Alternative names: DARPP 32 DARPP-32 Dopamine and cAMP regulated neuronal phosphoprotein 32 Dopamine and cAMP regulated neuronal phosphoprotein Dopamine and cAMP regulated phosphoprotein Dopamine and cAMP regulated phosphoprotein DARPP 32 Dopamine and cAMP regulated phosphoprotein DARPP32 Dopamine- and cAMP-regulated neuronal phosphoprotein FLJ20940 IPPD Neuronal phosphoprotein DARPP 32 PPP1R1B PPR1B_HUMAN Protein phosphatase 1 regulatory (inhibitor) subunit 1B Protein phosphatase 1 regulatory inhibitor subunit 1B Protein phosphatase 1 regulatory subunit 1B
Images
ET1608-23_1.jpg Fig1: Immunofluorescence analysis of frozen mouse striatum tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1608-23, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1608-23_2.jpg Fig2: Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1608-23, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1608-23_3.jpg Fig3: Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1608-23, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1608-23_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse forebrain tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/1,000 dilution for 1 hour 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.
ET1608-23_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse forebrain tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/1,000 dilution for 1 hour 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.
ET1608-23_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/500 dilution for 1 hour 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.
ET1608-23_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/500 dilution for 1 hour 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.
ET1608-23_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse brian tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/500 dilution for 1 hour 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.
ET1608-23_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/500 dilution for 1 hour 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.
ET1608-23_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat brian tissue with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-23) at 1/1,000 dilution for 1 hour 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.
ET1608-23_11.jpg Fig11: Western blot analysis of DARPP32 on different lysates with Rabbit anti-DARPP32 antibody (ET1608-23) at 1/2,000 dilution.

Lane 1: Human brain tissue lysate (40 µg/Lane)
Lane 2: Mouse brain tissue lysate (40 µg/Lane)
Lane 3: Rat brain tissue lysate (40 µg/Lane)
Lane 4: Rat hippocampus tissue lysate (40 µg/Lane)

Predicted band size: 23 kDa
Observed band size: 32 kDa

Exposure time: 12 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-23) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1608-23_12.jpg Fig12: 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).
ET1608-23_13.jpg Fig13: 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).
ET1608-23_14.jpg Fig14: 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).
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