Overview

  • Product name

    Anti-Neurogranin antibody [EPR21152]
    See all Neurogranin primary antibodies
  • Description

    Rabbit monoclonal [EPR21152] to Neurogranin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-Fr, WB, IHC-P, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Neurogranin aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q92686

  • Positive control

    • WB: Human fetal brain lysate and brain cortex lysate; mouse cerebral cortex lysate. Rat brain lysate. IHC-P: Human cerebellum tissue; mouse and rat cerebrum tissue. IHC-Fr: Mouse and rat hippocampus CA1 tissue. IP: Mouse cerebral cortex lysate. ICC/IF: Primary rat cortical neurons
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab217672 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 1/3000.

Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

WB 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 8 kDa).
IHC-P 1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.
ICC/IF 1/1500.

Target

  • Function

    Acts as a "third messenger" substrate of protein kinase C-mediated molecular cascades during synaptic development and remodeling. Binds to calmodulin in the absence of calcium.
  • Tissue specificity

    In the cerebral cortex, found in the cell bodies of neurons in layers II-VI, and in apical and basal dendrites of pyramidal neurons. Is not found in the dendrites in patients with Alzheimer disease.
  • Sequence similarities

    Belongs to the neurogranin family.
    Contains 1 collagen-like domain.
    Contains 1 IQ domain.
  • Information by UniProt
  • Database links

  • Alternative names

    • Protein kinase C substrate RC3 antibody
    • Calmodulin binding protein antibody
    • Hng antibody
    • NEUG(55-78) antibody
    • NEUG_HUMAN antibody
    • Neurogranin (protein kinase C substrate) antibody
    • Ng antibody
    • NRGN antibody
    • Protein kinase C substrate antibody
    • RC3 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Neurogranin with ab217672 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Mainly cytoplasmic staining on rat cerebrum (PMID: 26076492) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-Neurogranin antibody [EPR21152] (ab217672) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate at 10 µg
    Lane 2 : Human brain cortex lysate at 10 µg
    Lane 3 : Mouse cerebral cortex lysate at 20 µg
    Lane 4 : Rat brain lysate at 20 µg

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 8 kDa
    Observed band size: 15 kDa
    why is the actual band size different from the predicted?



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

    Exposure times:

    Lane 1: 32 seconds

    Lane 2: 101 seconds

    Lane 3: 85 seconds

    Lane 4: 3 minutes 

    The observed molecular mass is consistent with the literature (PMID 17505539).

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus CA1 tissue labeling Neurogranin with ab217672 at 1/3000 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Positive staining in the stratum pyramidal neurons of hippocampus CA1 on mouse brain (PMID: 15389631; 21516261) is observed. Counter stained with DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20)

  • Immunocytochemistry/ Immunofluorescence analysis of primary rat cortical neurons labeling Neurogranin with ab217672 at 1/1500. The cells were fixed with 4% paraformaldehyde containing 0.2% picric acid in 0.1M phosphate buffer, pH 6.9 for 20 minutes. Permeabilization was with 0.3% Triton-X 100 in PBS (PBSTx). ab150068 at 1/200 was used as the secondary antibody

    The rat cortical neurons were cultured for 29 days in vitro. They were either left untreated (Control) or treated beginning on the 10th day with 60ng/mL triiodothyronine (T3), to enhance neurogranin expression.

    The cells were visualized with an inverted microscope at 10X magnification (upper panels) or 20X magnification (lower panels).

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized  frozen rat hippocampus CA1 tissue labeling Neurogranin with ab217672 at 1/3000 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Positive staining in the stratum pyramidal neurons of hippocampus CA1 on rat brain (PMID: 15389631; 21516261) is observed. Counter stained with DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20)

  • Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Neurogranin with ab217672 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Mainly cytoplasmic staining on mouse cerebrum (PMID: 26076492) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Neurogranin with ab217672 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Mainly cytoplasmic but also weak nuclear staining on human cerebrum (PMID: 26076492; PMID: 21516261) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Neurogranin was immunoprecipitated from 0.35 mg mouse cerebral cortex lysate with ab217672 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217672 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution.

    Lane 1: Mouse cerebral cortex lysate 10 µg (Input).
    Lane 2: ab217672 IP in mouse cerebral cortex lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217672 in mouse cerebral cortex lysate (-).

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 15 seconds.

References

ab217672 has not yet been referenced specifically in any publications.

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