Anti-Neurogranin 抗体 [EPR21152]

• IHC-P

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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 ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurogranin antibody [EPR21152] (AB217672)

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling NRGN with ab217672 at 0.1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab217672 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Supplier Data

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 ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

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 ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-Fr

Supplier Data

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 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)

• ICC/IF

Lab

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). 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).

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

• IHC-Fr

Supplier Data

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 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)

• IP

Supplier Data

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) (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.

All lanes:

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

Predicted band size: 8 kDa

Observed band size: 15 kDa

false

• WB

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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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Lanes 3 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 8 kDa

Observed band size: 15 kDa

true