Recombinant
RabMAb

Recombinant Anti-nNOS (neuronal) antibody [EP1855Y] - Low endotoxin, Azide free (ab219373)

Overview

  • Product name
    Anti-nNOS (neuronal) antibody [EP1855Y] - Low endotoxin, Azide free
    See all nNOS (neuronal) primary antibodies
  • Description
    Rabbit monoclonal [EP1855Y] to nNOS (neuronal) - Low endotoxin, Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, ICC/IF, IHC-Frmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Common marmoset
  • 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

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab219373 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 Use at an assay dependent concentration. Predicted molecular weight: 161 kDa.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • 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.
    • Tissue specificity
      Isoform 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 similarities
      Belongs to the NOS family.
      Contains 1 FAD-binding FR-type domain.
      Contains 1 flavodoxin-like domain.
      Contains 1 PDZ (DHR) domain.
    • Domain
      The 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 localization
      Cell 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

    Images

    • 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

    References

    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

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