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Rabbit Recombinant Multiclonal nNOS antibody. Suitable for IHC-P, WB, IHC-Fr, ICC/IF and reacts with Transfected cell line - Human, Human, Mouse, Rat samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (AB323183), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (AB323183), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (AB323183), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (AB323183), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (AB323183), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Multiclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBIHC-FrICC/IFIPFlow Cyt (Intra)
Human
Tested
Tested
Expected
Not recommended
Not recommended
Not recommended
Mouse
Tested
Tested
Tested
Tested
Not recommended
Not recommended
Rat
Tested
Tested
Tested
Tested
Not recommended
Not recommended
Transfected cell line - Human
Tested
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Transfected cell line - Human

Dilution info
1/5000
Notes

-

Species

Human

Dilution info
1/100
Notes

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

Mouse

Dilution info
1/50
Notes

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

Rat

Dilution info
1/50
Notes

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

Tested
Tested

Species

Human

Dilution info
1/1000
Notes

-

Species

Mouse

Dilution info
1/1000
Notes

-

Species

Rat

Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info
1/100
Notes

-

Species

Rat

Dilution info
1/100
Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Transfected cell line - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info
1/2000
Notes

-

Species

Rat

Dilution info
1/2000
Notes

-

Not recommended
Not recommended

Species

Human, Transfected cell line - Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat, Transfected cell line - Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat, Transfected cell line - Human

Dilution info

-

Notes

-

Target data

Function

Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In the brain and peripheral nervous system, NO displays many properties of a neurotransmitter. Probably has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such SRR.

Targets

Nos1, Nos1

Alternative names

Rabbit Recombinant Multiclonal nNOS antibody. Suitable for IHC-P, WB, IHC-Fr, ICC/IF and reacts with Transfected cell line - Human, Human, Mouse, Rat samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Multiclonal

Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

RM1237

Purification technique

Affinity purification Protein A

Specificity

Unsuitable for human ICC.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

19 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on human cerebellum. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on myenteric nerve plexus of human colon. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling nNOS (neuronal) with ab323183 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on mouse cerebrum. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling nNOS (neuronal) with ab323183 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on rat cerebrum. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Low expression: no staining on human spleen. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling nNOS (neuronal) with ab323183 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Low expression: no staining on mouse spleen. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling nNOS (neuronal) with ab323183 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Low expression: no staining on rat spleen. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human NOS1 expression vector containing a myc-His-tag cell pellet. (B) HEK-293T transfected with a human NOS2 expression vector containing a myc-His-tag cell pellet. (C) HEK-293T transfected with a human NOS3 expression vector containing a myc-His-tag cell pellet. (D) HEK-293T transfected with an empty vector containing a His-tag. tissue labeling nNOS (neuronal) with ab323183 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on (A) HEK-293T transfected with a human NOS1 expression vector containing a myc-His-tag cell pellet, no staining on (B) HEK-293T transfected with a human NOS2 expression vector containing a myc-His-tag cell pellet, (C) HEK-293T transfected with a human NOS3 expression vector containing a myc-His-tag cell pellet, and (D) HEK-293T transfected with an empty vector containing a His-tag cell pellet. The section was incubated with ab323183 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

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

    Low expression tissue: spleen

    The identity of the lower MW band at approximately 100 kDa (in lane 1, 2, 4, 5) is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183) at 1/1000 dilution

    Lane 1: Mouse cerebellum tissue lysate at 20 µg

    Lane 2: Mouse brain tissue lysate at 20 µg

    Lane 3: Mouse spleen tissue lysate at 20 µg

    Lane 4: Rat cerebellum tissue lysate at 20 µg

    Lane 5: Rat brain tissue lysate at 20 µg

    Lane 6: Rat spleen tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 161 kDa, 36 kDa

    Exposure time: 10s

  • Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 4T1(mouse mammary gland carcinoma epithelial cell) cells labelling nNOS (neuronal) with ab323183 at 1/2000 (0.253 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

    Low expression: Confocal image showing no staining in 4T1 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cells labelling nNOS (neuronal) with ab323183 at 1/2000 (0.253 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

    Confocal image showing cytoplasmic staining in rat primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled 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).

    Anti-MAP2 antibody [HM-2] ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4ug/ml) dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2ug/ml) dilution (Magenta).

    -ve control 1: ab323183 at a 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at a 1/1000 dilution.

    -ve control 2: Anti-MAP2 antibody [HM-2] ab11267 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution

  • Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labelling nNOS (neuronal) with ab323183 at 1/2000 (0.253 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

    Confocal image showing cytoplasmic staining in mouse primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled 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).

    Anti-MAP2 antibody [HM-2] ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4ug/ml) dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2ug/ml) dilution (Magenta).

    -ve control 1: ab323183 at a 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at a 1/1000 dilution.

    -ve control 2: Anti-MAP2 antibody [HM-2] ab11267 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution

  • Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh frozen) tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

    Low expression: Confocal image showing no staining on mouse spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323183 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

  • Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (fresh frozen) tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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-nNOS (neuronal) (ab323183, green), anti-Synapsin I (Alexa Fluor® 594 Anti-Synapsin I - Synaptic Marker antibody [EPR23531-50] ab309972, magenta) and anti-NeuN (Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, ) on rat cerebellum.

    Panel B: anti-nNOS (neuronal) stained on rat cerebellum.

    Panel C: anti-Synapsin I stained in neurons of rat cerebellum.

    Panel D: anti-NeuN stained in neurons of rat cerebellum.

    The section was incubated in two rounds of staining: in the order of ab323183 and Alexa Fluor® 594 Anti-Synapsin I - Synaptic Marker antibody [EPR23531-50] ab309972, Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565 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).

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

  • Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh frozen) tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

    Low expression: Confocal image showing no staining on rat spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323183 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

  • Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh frozen) tissue labeling nNOS (neuronal) with ab323183 at 1/100 (5.05 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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-nNOS (neuronal) (ab323183, green), anti-Synapsin I (Alexa Fluor® 594 Anti-Synapsin I - Synaptic Marker antibody [EPR23531-50] ab309972, magenta) and anti-NeuN (Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565, ) on mouse cerebellum.

    Panel B: anti-nNOS (neuronal) stained on mouse cerebellum.

    Panel C: anti-Synapsin I stained in neurons of mouse cerebellum.

    Panel D: anti-NeuN stained in neurons of mouse cerebellum.

    The section was incubated in two rounds of staining: in the order of ab323183 and Alexa Fluor® 594 Anti-Synapsin I - Synaptic Marker antibody [EPR23531-50] ab309972, Alexa Fluor® 647 Anti-NeuN antibody [EPR12763] - Neuronal Marker ab190565 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).

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

  • Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

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

    Low expression tissue: spleen, pancreas

    The identity of the lower MW band at approximately 100 kDa (in lane 1) is unknown.

    In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) staining at 1/100000 dilution (15 kDa).

    All lanes: Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183) at 1/1000 dilution

    Lane 1: Human hippocampus tissue lysate at 20 µg

    Lane 2: Human spleen tissue lysate at 20 µg

    Lane 3: Human pancreas tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 161 kDa, 15 kDa

    Exposure time: 37s

  • Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

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

    Negative control: HeLa

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183) at 1/1000 dilution

    Lane 1: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 161 kDa, 36 kDa

    Exposure time: 180s

  • Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183), expandable thumbnail

    Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183)

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

    Low expression: 4T1

    This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    All lanes: Western blot - Anti-nNOS (neuronal) antibody [RM1237] (ab323183) at 1/1000 dilution

    Lane 1: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

    Lane 2: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: 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.

    Observed band size: 161 kDa, 124 kDa

    Exposure time: 180s

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