Recombinant
RabMAb

Anti-DARPP32 antibody [EP720Y] (ab40801)

Overview

  • Product name
    Anti-DARPP32 antibody [EP720Y]
    See all DARPP32 primary antibodies
  • Description
    Rabbit monoclonal [EP720Y] to DARPP32
  • Tested applications
    Suitable for: 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-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 protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

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

  • 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

  • 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. 

  • 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.

  • 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.

  • 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
    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 as the secondary antibody (1/1000).

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

     

     

  • 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.

  • 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.

  • 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. 

References

This product has been referenced in:
  • Kosubek-Langer J  et al. Maturation, Behavioral Activation, and Connectivity of Adult-Born Medium Spiny Neurons in a Striatal Song Nucleus. Front Neurosci 11:323 (2017). IHC ; Zebra finch . Read more (PubMed: 28638318) »
  • Straccia M  et al. Human t-DARPP is induced during striatal development. Neuroscience 333:320-30 (2016). WB, IHC-P ; Human . Read more (PubMed: 27475250) »

See all 25 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Treatment
alpha synuclein
Specification
human
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Ms. Sehee Oh

Verified customer

Submitted Jul 13 2016

Application
Flow Cytometry
Sample
Human Cell (Differentiated hNSCs)
Permeabilization
Yes - 0.25% Triton X-100 in DPBS
Gating Strategy
Undifferentiated Stem Cells (white)
Specification
Differentiated hNSCs
Preparation
Cell harvesting/tissue preparation method: Accutase
Sample buffer: PBS
Fixation
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 18 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Human Cell (Differentiated hNSCs)
Specification
Differentiated hNSCs
Permeabilization
Yes - 0.25% Triton X-100 in DPBS
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 17 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Sheep Tissue sections (Brain Median Eminence)
Specification
Brain Median Eminence
Permeabilization
Yes - 0.01% Triton
Fixative
Paraformaldehyde
Username

Dr. Gregroy Conductier

Verified customer

Submitted Oct 24 2014

Application
IHC - Wholemount
Sample
Zebra finch Tissue (14um brain section)
Specification
14um brain section
Username

Abcam user community

Verified customer

Submitted May 13 2014

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (12% Tris Glycine)
Sample
Zebra finch Tissue lysate - other (Brain cytoplasmic fraction)
Specification
Brain cytoplasmic fraction
Blocking step
Rotiblock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 14 2013

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 ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - other (Human brain tissue homogenate)
Loading amount
5 µg
Specification
Human brain tissue homogenate
Treatment
Sample boiled in sample buffer for 5 mins prior to loading on gel
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Aug 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Pig Tissue sections (Brain)
Specification
Brain
Fixative
Formaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 0.2% · Temperature: 24°C
Username

Mr. Marian Hruska-Plochan

Verified customer

Submitted Feb 24 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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