Anti-Neuroligin 2 antibody [EPR28230-122]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse NLGN2 expression vector containing a his tag. (B) HEK-293T cells transfected with a mouse NLGN1 expression vector containing a his tag. (C) HEK-293T cells transfected with a mouse NLGN3 expression vector containing a his tag. (D) HEK-293T cells transfected with empty vector containing a his tag. tissue labeling Neuroligin 2 with ab317303 at 1/100 (4.92 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T cells transfected with a mouse NLGN2 expression vector containing a his tag, no staining on (B) HEK-293T cells transfected with a mouse NLGN1 expression vector containing a his tag, and (C) HEK-293T cells transfected with a mouse NLGN3 expression vector containing a his tag, and (D) HEK-293T cells transfected with empty vector containing a his tag.
The section was incubated with ab317303 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Neuroligin 2 with ab317303 at 1/100 (4.92 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat liver.
The section was incubated with ab317303 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Neuroligin 2 with ab317303 at 1/100 (4.92 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver.
The section was incubated with ab317303 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh frozen) tissue labeling Neuroligin 2 with ab317303 at 1/50 (9.84 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Negative control : confocal image showing no staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317303 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

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Immunohistochemistry (Frozen sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling Neuroligin 2 with ab317303 at 1/50 (9.84 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Negative control : confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317303 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• Flow Cyt

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Flow Cytometry - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Flow cytometric analysis of Rat primary neuron cells labelling Neuroligin 2 with ab317303 at 1/50 dilution (1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cell.

• Flow Cyt

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Flow Cytometry - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Flow cytometric analysis of Mouse primary neuron cells labelling Neuroligin 2 with ab317303 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cell.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh frozen) tissue labeling Neuroligin 2 with ab317303 at 1/50 (9.84 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Panel A : merged staining of anti-Neuroligin-2 (ab317303, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on rat hippocampus.
Panel B : anti-Neuroligin-2 stained on rat hippocampus.
Panel C : anti-NeuN stained in neurons of rat hippocampus.
Panel D : anti-GFAP stained in astrocytes of rat hippocampus.
The section was incubated in two rounds of staining : in the order of ab317303 and ab190565, ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling Neuroligin 2 with ab317303 at 1/50 (9.84 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling Neuroligin 2 with ab317303 at 1/50 (9.84 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Panel A : merged staining of anti-Neuroligin-2 (ab317303, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on mouse hippocampus.
Panel B : anti-Neuroligin-2 stained on mouse hippocampus.
Panel C : anti-NeuN stained in neurons of mouse hippocampus.
Panel D : anti-GFAP stained in astrocytes of mouse hippocampus.
The section was incubated in two rounds of staining : in the order of ab317303 and ab190565, ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Neuroligin 2 with ab317303 at 1/100 (4.92 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum (PMID : 16982420).
The section was incubated with ab317303 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Neuroligin 2 with ab317303 at 1/100 (4.92 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum (PMID : 19926856).
The section was incubated with ab317303 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-Neuroligin 2 antibody [EPR28230-122] (AB317303)

Neuroligin 2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab317303 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317303 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate
Lane 2 : ab317303 IP in Mouse brain tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317303 in mouse brain tissue lysate

All lanes:

Immunoprecipitation - Anti-Neuroligin 2 antibody [EPR28230-122] (ab317303) at 1/30 dilution

All lanes:

Mouse brain tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s