Overview

  • Product name
    Anti-DARPP32 antibody [EP720Y]
    See all DARPP32 primary antibodies
  • Description
    Rabbit monoclonal [EP720Y] to DARPP32
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC - Wholemount, IHC-Fr, Flow Cyt, ICC/IF, WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic peptide within Human DARPP32 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q9UD71
    (Peptide available as ab189245)

  • Positive control
    • Rat brain, cerebral cortex and hippocampus tissue; Mouse brain and cerebral cortex tissue; Human breast adenocarcinoma; Human fetal brain tissue lysate; Human colon tissue; pig brain tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab40801 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC - Wholemount Use at an assay dependent concentration. PubMed: 21394212
IHC-Fr 1/7500.
Flow Cyt Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/50 - 1/100.
WB 1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).Can be blocked with DARPP32 peptide (ab189245).

For unpurified use at 1/5000 - 1/50 000. 

IHC-P 1/50.

See IHC antigen retrieval protocols.

IP 1/20.

For unpurified use at 1/50. 

Target

  • Function
    Inhibitor of protein-phosphatase 1.
  • Sequence similarities
    Belongs to the protein phosphatase inhibitor 1 family.
  • Post-translational
    modifications
    Dopamine- and cyclic AMP-regulated neuronal phosphoprotein.
    Phosphorylation of Thr-34 is required for activity.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human colon sections labelling DARPP32 with purified ab40801 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence analysis of mouse brain tissue lysate labelling DARPP32 with purified ab40801 at 1/100 (3.4 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was performed with Heated citrate solution (10mM citrate PH 6.0 + 0.05% Tween-20). ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody.

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control and is shown in the inset. 

  • All lanes : Anti-DARPP32 antibody [EP720Y] (ab40801) at 1/50000 dilution (purified)

    Lane 1 : Rat Hippocampus tissue lysate
    Lane 2 : Rat brain tissue lysate
    Lane 3 : Mouse cerebral cortex tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

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



    Blocking/Diluting buffer 5% NFDM/TBST
  • ab40801 at 1/20 immunoprecipitating DARPP32 in rat brain whole cell lysate observed at 32 KDa (lanes 1 and 2).

    Lane 1 (input): Rat brain whole cell lysate 10μg

    Lane 2 (+): ab40801 + Rat brain whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40801 in Rat brain whole cell lysate

    For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

     

     

  • Immunohistochemical analysis of murine brain tissue, staining DARPP32 (red) with unpurified ab40801. Slices were fixed in paraformaldehyde and blocked with 5% horse serum and 0.25% Triton X-100. Samples were incubated with primary antibody (1/200) for 15 hours.

  • ab40801 (unpurified) staining DARPP32 in pig brain tissue sections by IHC-Fr (formaldehyde-fixed Frozen sections). Tissue was fixed with formaldehyde (4%), permeabilized with Triton X-100 and blocked with 0.2% milk for 30 minutes at 24°C. Samples were incubated with primary antibody (1/7500 in TBS + 1% Triton X-100 + 0.2% milk) for 24 hours at 4°C. A Biotin-conjugated Sheep polyclonal to rabbit IgG, dilution 1/400, was used as secondary antibody. Endogenous peroxidase was blocked with 0.3% H2O2 in methanol for 20 minutes.

    See Abreview

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex sections labelling DARPP32 with purified ab40801 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Anti-DARPP32 antibody [EP720Y] (ab40801) at 1/1000 dilution (purified) + Human fetal brain lysate at 15 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

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



    Blocking/Diluting buffer 5% NFDM/TBST
  • Anti-DARPP32 antibody [EP720Y] (ab40801) at 1/50000 dilution (unpurified) + Rat brain lysate at 10 µg

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

  • Immunohistochemical analysis of human breast adenocarcinoma sections labelling DARPP32 with unpurified ab40801 at a dilution of 1/50. 

  • Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex sections labelling DARPP32 with purified ab40801 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

References

This product has been referenced in:
  • Shi Z  et al. miR-9 regulates basal ganglia-dependent developmental vocal learning and adult vocal performance in songbirds. Elife 7:N/A (2018). Read more (PubMed: 29345619) »
  • Weber JJ  et al. Calpastatin ablation aggravates the molecular phenotype in cell and animal models of Huntington disease. Neuropharmacology 133:94-106 (2018). WB . Read more (PubMed: 29355642) »
See all 41 Publications for this product

Customer reviews and Q&As

Answer

Thank you for contacting us.

While there a many targets which express differentially in distinct layers of the neocortex many of these are expressed in the striatum as well. I found these papers to have a number of good targets which seem specific, such as Pax6 or, for striatal markers, DARPP32, CalB or GAD67. However a search through the Allen Mouse Brain Atlas expression data shows similar gene expression of all these markers in adult striatum and cortex. I would emphasize that this is not vetted data.

While any single or combination of these markers may well yield the information which you will be looking for, I feel that the best choices may be to try a few of these targets presented within this literature as you would have reference material to compare with.

Further review of our catalog leads me to suggest use of a DARPP32 antibody such as ab40801 alongside use of a Vementin antibody such as ab8978. Alternatively use of PAX6 ab5790 alongside GAD67, ab26116, may yield good results.

Each of these pairs will work in double staining as they are comprised of a mouse and a rabbit derived antibody. Further each product has been used in numerous publications and is well characterized by these, Abreviews and scientific support on the datasheet. Each is guaranteed to work in IHC-P as well as a number of other applications and would therefore be covered by our Abpromise guarantee:https://www.abcam.com/abpromise.

I have added links to each of these products below for your review:

ab40801;https://www.abcam.com/DARPP32-antibody-EP720Y-ab40801.html

ab8978;https://www.abcam.com/Vimentin-antibody-RV202-Neural-Stem-Cell-Marker-ab8978.html

ab5790;https://www.abcam.com/PAX6-antibody-Stem-Cell-Marker-ab5790.html

ab26116;https://www.abcam.com/GAD67-antibody-K-87-Neuronal-Marker-ab26116.html



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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