Anti-Neurogranin antibody [EPR21152]

8 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Western blot - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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.

  All lanes: Western blot - 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: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Lanes 3 - 4: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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

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  Immunohistochemistry (Frozen sections) - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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)

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  Image courtesy of Ms. Babben Tinner, QBM Cell Science.

  Immunocytochemistry/ Immunofluorescence - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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). Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed 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 10^th 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).

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  Immunohistochemistry (Frozen sections) - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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)

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  Immunoprecipitation - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab217672 in mouse cerebral cortex lysate (-).

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

  All lanes: Immunoprecipitation - Anti-Neurogranin antibody [EPR21152] (ab217672)

  Predicted band size: 8 kDa

  Observed band size: 15 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurogranin antibody [EPR21152] (ab217672)

  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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).