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RecombinantRabMAb

Recombinant Anti-DARPP32 antibody [EP720Y] (ab40801)

  • Datasheet
  • SDS
Reviews (9)Q&A (1)References (60)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Immunocytochemistry/ Immunofluorescence - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Immunoprecipitation - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
  • Anti-DARPP32 antibody [EP720Y] (ab40801)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP720Y] to DARPP32
  • Suitable for: ICC/IF, WB, IHC-P, IP
  • Reacts with: Mouse, Rat, Human

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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: ICC/IF, WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: 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

    • IHC-P: Human breast adenocarcinoma tissue and colon tissue. Mouse and rat cerebral cortex tissue. ICC/IF: Mouse brain tissue. WB: Rat hippocampus and brain tissue lysate. Mouse cerebral cortex tissue lysate. Human fetal brain lysate. IP: Rat brain whole cell lysate. IHC-Wm: Mouse brain tissue. IHC-Fr: Pig brain tissue.
  • General notes

    Protein phosphatase 1 regulatory subunit 1B (PPP1R1B) is a protein that in humans is encoded by the PPP1R1B gene. This gene, also known as DARPP-32, has a role as a dopamine- and cyclic AMP-regulated phosphoprotein. As such PPP1R1B affects dopamine glutamate and adenosine; and potentially has a role in conditions such as schizophrenia and Parkinson's disease. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP720Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Receptors / Channels
    • GPCR
    • Dopamine Receptors
    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Metabolism
    • Types of disease
    • Obesity

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-DARPP32 antibody [EP720Y] (ab199776)
    • Anti-DARPP32 antibody [EP720Y] - BSA and Azide free (ab220808)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Rat brain cerebrum tissue lysate - total protein (ab4033)

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
ICC/IF 1/50 - 1/100.
WB 1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).

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

IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession Q9UD71 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 84152 Human
    • Entrez Gene: 19049 Mouse
    • Entrez Gene: 360616 Rat
    • Omim: 604399 Human
    • SwissProt: Q9UD71 Human
    • SwissProt: Q60829 Mouse
    • SwissProt: Q6J4I0 Rat
    • Unigene: 286192 Human
    • Unigene: 45372 Mouse
    • Unigene: 70366 Rat
    see all
  • 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)

    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.

  • Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Immunocytochemistry/ Immunofluorescence - Anti-DARPP32 antibody [EP720Y] (ab40801)

    Immunocytochemistry/Immunofluorescence analysis of mouse brain tissue 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. 

  • Immunoprecipitation - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Immunoprecipitation - Anti-DARPP32 antibody [EP720Y] (ab40801)

    ab40801 at 1/20 immunoprecipitating DARPP32 in rat brain whole cell lysate observed at 32 KDa (lanes 1 and 2).

    Lane 1: Rat brain whole cell lysate 10μg.

    Lane 2: ab40801 IP in 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/Dilution buffer and concentration: 5% NFDM/TBST.

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)

    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.

  • Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
    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
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)

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

  • Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801)
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (ab40801)

    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)
    Anti-DARPP32 antibody [EP720Y] (ab40801)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (60)

    Publishing research using ab40801? Please let us know so that we can cite the reference in this datasheet.

    ab40801 has been referenced in 60 publications.

    • Li D  et al. Gastro-Protective Effects of Calycosin Against Precancerous Lesions of Gastric Carcinoma in Rats. Drug Des Devel Ther 14:2207-2219 (2020). PubMed: 32606591
    • Liu Q  et al. Cerebellum-enriched protein INPP5A contributes to selective neuropathology in mouse model of spinocerebellar ataxias type 17. Nat Commun 11:1101 (2020). PubMed: 32107387
    • Nogi Y  et al. Expression of feeding-related neuromodulatory signalling molecules in the mouse central olfactory system. Sci Rep 10:890 (2020). PubMed: 31964903
    • Williams GP  et al. T cell infiltration in both human multiple system atrophy and a novel mouse model of the disease. Acta Neuropathol 139:855-874 (2020). PubMed: 31993745
    • Zhao Y  et al. ATAD3A oligomerization causes neurodegeneration by coupling mitochondrial fragmentation and bioenergetics defects. Nat Commun 10:1371 (2019). PubMed: 30914652
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-10 of 10 Abreviews or Q&A

    Immunohistochemistry free floating abreview for Anti-DARPP32 antibody [EP720Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (Brain, 45 µm VIBRATOME sections)
    Specification
    Brain, 45 µm VIBRATOME sections
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 07 2019

    Western blot abreview for Anti-DARPP32 antibody [EP720Y]

    Good
    Abreviews
    Abreviews
    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
    Read More

    Ms. Sehee Oh

    Verified customer

    Submitted Jul 13 2016

    Flow Cytometry abreview for Anti-DARPP32 antibody [EP720Y]

    Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 18 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-DARPP32 antibody [EP720Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 17 2015

    Immunohistochemistry (Frozen sections) abreview for Anti-DARPP32 antibody [EP720Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Dr. Gregroy Conductier

    Verified customer

    Submitted Oct 24 2014

    IHC - Wholemount abreview for Anti-DARPP32 antibody [EP720Y]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Zebra finch Tissue (14um brain section)
    Specification
    14um brain section
    Read More

    Abcam user community

    Verified customer

    Submitted May 13 2014

    Western blot abreview for Anti-DARPP32 antibody [EP720Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 14 2013

    Question

      Thank you so much for helping me find primary (and secondary) antibodies for IHC on mouse neural progenitors down the lineage of cortex verses striatum. Attached is the paper (2012) that mentions cortex being identified with NCAM (Figure 4 i,j page 481) and Figure 1 says cortical induction efficiency was measured by Pax6, Vimentin, Otx1 and Otx2 expression (Figure 1c, page 478).   The second paper I attached (2008) is to help us figure out a good primary/secondary antibody detection of striatal differentiation. It says “Terminal striatal differentiation can be established by the detection ofMAP-2/Ki67[1] postmitotic neurons expressing key striatal markers, such as GABA, GAD67 (GAD1), DARPP32, ARPP21, calbindin, or calretinin.”   If you can find anything more efficient and specific for cortex vs. striatum it would be wonderful! Please contact me for any reason and thank you for your time and energy,

    Read More

    Abcam community

    Verified customer

    Asked on May 23 2012

    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.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    Abcam Scientific Support

    Answered on May 23 2012

    Western blot abreview for Anti-DARPP32 antibody [EP720Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 19 2011

    Immunohistochemistry (Frozen sections) abreview for Anti-DARPP32 antibody [EP720Y]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mr. Marian Hruska-Plochan

    Verified customer

    Submitted Feb 24 2010

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