Anti-nNOS (neuronal) antibody [EP1855Y] (ab76067)

Overview

  • Product nameAnti-nNOS (neuronal) antibody [EP1855Y]
    See all nNOS (neuronal) primary antibodies
  • Description
    Rabbit monoclonal [EP1855Y] to nNOS (neuronal)
  • Tested applicationsSuitable for: WB, IP, Flow Cyt, IHC-Fr, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Marmoset (common)
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human nNOS (neuronal). A synthetic peptide corresponding to residues around serine 1417 of human nNOS (neuronal) protein.

  • Positive control
    • WB: Mouse brain tissue lysate. ICC/IF: A673 cells. Flow Cyt: PC-12 cells. IP: Rat brain tissue lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:


    Anti-nNOS (neuronal) antibody (Alexa Fluor® 594) [EP1855Y] (ab198516)

    Anti-nNOS (neuronal) antibody (Alexa Fluor® 647) [EP1855Y] (ab198327)

Properties

Applications

Our Abpromise guarantee covers the use of ab76067 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
WB 1/1000 - 1/2000. Predicted molecular weight: 161 kDa.

For unpurified use at 1/100.

IP 1/150.

For unpurified use at 1/4.

Flow Cyt 1/600.

For unpurified use at 1/15.





ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr 1/1000.
ICC/IF 1/200.
  • Application notesIs unsuitable for IHC-P.
  • Target

    • FunctionProduces 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.
    • Tissue specificityIsoform 1 is ubiquitously expressed: detected in skeletal muscle and brain, also in testis, lung and kidney, and at low levels in heart, adrenal gland and retina. Not detected in the platelets. Isoform 3 is expressed only in testis. Isoform 4 is detected in testis, skeletal muscle, lung, and kidney, at low levels in the brain, but not in the heart and adrenal gland.
    • Sequence similaritiesBelongs to the NOS family.
      Contains 1 FAD-binding FR-type domain.
      Contains 1 flavodoxin-like domain.
      Contains 1 PDZ (DHR) domain.
    • DomainThe PDZ domain in the N-terminal part of the neuronal isoform participates in protein-protein interaction, and is responsible for targeting nNos to synaptic membranes in muscles. Mediates interaction with VAC14.
    • Post-translational
      modifications
      Ubiquitinated; mediated by STUB1/CHIP in the presence of Hsp70 and Hsp40 (in vitro).
    • Cellular localizationCell membrane > sarcolemma. Cell projection > dendritic spine. In skeletal muscle, it is localized beneath the sarcolemma of fast-twitch muscle fiber by associating with the dystrophin glycoprotein complex. In neurons, enriched in dendritic spines.
    • Information by UniProt
    • Database links
    • Alternative names
      • 2310005C01Rik antibody
      • BNOS antibody
      • Constitutive NOS antibody
      • EC 1.14.13.39 antibody
      • IHPS 1 antibody
      • IHPS1 antibody
      • N-NOS antibody
      • NC-NOS antibody
      • neuronal Nitric Oxide Synthase antibody
      • Neuronal NOS antibody
      • Nitric oxide synthase , neuronal, included antibody
      • Nitric oxide synthase 1 (neuronal) antibody
      • Nitric oxide synthase 1 antibody
      • Nitric oxide synthase, brain antibody
      • Nitric oxide synthase, penile neuronal, included antibody
      • NNOS antibody
      • NO antibody
      • NOS 1 antibody
      • NOS antibody
      • NOS type I antibody
      • NOS-I antibody
      • NOS1 antibody
      • NOS1_HUMAN antibody
      • Peptidyl-cysteine S-nitrosylase NOS1 antibody
      see all

    Anti-nNOS (neuronal) antibody [EP1855Y] images

    • Anti-nNOS (neuronal) antibody [EP1855Y] (ab76067) at 1/100 dilution (unpurified) + Mouse brain tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 161 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-nNOS (neuronal) antibody [EP1855Y] (ab76067) at 1/3000 dilution (purified) + Mouse brain tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 161 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-nNOS (neuronal) antibody [EP1855Y] (ab76067) at 1/1000 dilution (unpurified) + Mouse brain tissue lysate at 10 µg

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size : 161 kDa
      Observed band size : 161 kDa
    • Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling nNOS (neuronal) (green) with purified ab76067 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/200) and secondary antibody Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    • Unpurified ab76067 staining nNOS (neuronal) in murine sperm cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde and blocked using 2% BSA. Samples were then incubated with undiluted ab76067. The secondary used was a FITC conjugated goat anti-rabbit IgG at a 1/400 dilution.Panel A shows the specific staining of nNOS in sperm while Panel B is the control sample treated with Rabbit IgG.

      See Abreview

    • Overlay histogram showing PC-12 cells stained with unpurified ab76067 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (unpurified ab76067, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in PC-12 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.
      Please note that Abcam do not have any data for use of this antibody in non-fixed cells. We welcome any customer feedback.

    • Flow cytometry analysis of PC-12 cells labelling nNos (neuronal) with unpurified ab76067 at 1/15 (red). Cells were fixed with 100% methanol. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

    • Flow cytometry analysis of PC-12 cells labelling nNos (neuronal) with purified ab76067 at 1/600 (red). Cells were fixed with 100% methanol. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

    • ab76067 (unpurified) at 1/4 immunoprecipitating nNOS (neuronal) in rat brain tissue lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • ab76067 (purified) at 1/150 immunoprecipitating nNOS (neuronal) in rat brain tissue lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • IHC-Fr image of nNOS staining on marmoset caudate sections using unpurified ab76067 (1/1000). The tissue was fixed in paraformaldehyde and the sections were then permeabilized using Triton-X. The sections were them blocked using 2% BSA for 2 hour at 20°C. Unpurified ab76067 was diluted 1/1000 and incubated with the sections for 18 hours at 20°C. The secondary antibody used was HRP conjugated goat polyclonal to rabbit IgG (1/1000).

      See Abreview

    References for Anti-nNOS (neuronal) antibody [EP1855Y] (ab76067)

    This product has been referenced in:
    • Nishimatsu H  et al. Senescent Cells Impair Erectile Function through Induction of Endothelial Dysfunction and Nerve Injury in Mice. PLoS One 10:e0124129 (2015). IHC . Read more (PubMed: 25894557) »
    • Liu Q  et al. Melanoma NOS1 expression promotes dysfunctional IFN signaling. J Clin Invest 124:2147-59 (2014). Flow Cyt ; Human . Read more (PubMed: 24691438) »

    See all 12 Publications for this product

    Product Wall

    Application Immunohistochemistry free floating
    Sample Mouse Tissue sections (Brain)
    Specification Brain
    Username

    Dr. Hyun Jin Kim

    Verified customer

    Submitted Mar 21 2016

    Application Western blot
    Sample Rat Tissue lysate - whole (Skeletal muscle)
    Gel Running Conditions Reduced Denaturing (8%)
    Loading amount 20 µg
    Specification Skeletal muscle
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 09 2016

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (8%)
    Sample Rat Tissue lysate - whole (Skeletal muscle)
    Specification Skeletal muscle
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 02 2015

    Application Immunohistochemistry (Frozen sections)
    Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
    Sample Marmoset (common) Tissue sections (Caudate)
    Specification Caudate
    Permeabilization Yes - Triton-X
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Feb 21 2014

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (8% Gel)
    Sample Human Tissue lysate - whole (vastus lateralis muscle)
    Specification vastus lateralis muscle
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 18 2014

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (8)
    Sample Mouse Tissue lysate - whole (Skeletal muscle)
    Specification Skeletal muscle
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 09 2014


    The NOS1 protein of Mustela is 95% similar to Bovine nNOS so it seems like the antibody ab2801 could be an option. It suppose to detect brain Nitric Oxide Synthase.

    As Ferret is an untested species so in case you are interested in buying ...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Tissue lysate - whole (Skeletal muscle)
    Loading amount 20 µg
    Specification Skeletal muscle
    Gel Running Conditions Reduced Denaturing (8% Gel)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Dr. G.K Sakellariou

    Verified customer

    Submitted May 14 2013

    Vielen Dank für Ihre Antwort.

    Die Referenznummer Ihrer Gutschrift ist 123456.

    Es tut mir leid, dass der Antikörper nicht so funktioniert hat, wie auf dem Datenblatt angegeben. Ich habe unsere Finanzabteilung beauftragt, ...

    Read More

    Vielen Dank für Ihre Email.

    Frau Hoffmann ist nun in Ihrem wohlverdienten Urlaub, daher werde ich mich um Ihren Fall kümmern.

    Können Sie mir bitte mitteilen, wann Sie den Ersatz von uns erhalten haben, damit ich Ihne...

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