Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)
Key features and details
- Rabbit polyclonal to nNOS (neuronal) (phospho S847)
- Suitable for: IHC-FoFr, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-nNOS (neuronal) (phospho S847) antibody
See all nNOS (neuronal) primary antibodies -
Description
Rabbit polyclonal to nNOS (neuronal) (phospho S847) -
Host species
Rabbit -
Specificity
This antibody shows a reduction in signal when blocked with unmodified nNOS (neuronal) peptide in WB, however when tested in ELISA, it showed less than 2% cross reactivity with the unmodified protein. -
Tested applications
Suitable for: IHC-FoFr, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Rabbit, Xenopus laevis, Zebrafish, Apteronotus leptorhynchus
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Mouse nNOS (neuronal), phosphorylated at S847.
Read Abcam's proprietary immunogen policy (Peptide available as ab16981.)
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab16650 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 IHC-FoFr 1/3000. WB Use a concentration of 1 µg/ml. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa). Target
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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
modificationsUbiquitinated; 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
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Database links
- Entrez Gene: 4842 Human
- Entrez Gene: 18125 Mouse
- Entrez Gene: 100009243 Rabbit
- Entrez Gene: 24598 Rat
- Entrez Gene: 60658 Zebrafish
- Omim: 163731 Human
- SwissProt: P29475 Human
- SwissProt: Q9Z0J4 Mouse
see all -
Alternative names
- 2310005C01Rik antibody
- BNOS antibody
- Constitutive NOS antibody
see all
Images
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Western blot - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)All lanes : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml
Lane 1 : Forebrain (Mouse) Tissue Lysate
Lane 2 : Spinal Cord (Mouse) Tissue Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control)
Lane 4 : Forebrain (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide at 1 µg/ml
Lane 5 : Spinal Cord (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide at 1 µg/ml
Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control) with Mouse nNOS (neuronal) (phospho S847) peptide at 1 µg/ml
Lane 7 : Forebrain (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (unmodified) peptide at 1 µg/ml
Lane 8 : Spinal Cord (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (unmodified) peptide at 1 µg/ml
Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control) with Mouse nNOS (neuronal) (unmodified) peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutesThis blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab16650 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Western blot - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)All lanes : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml
Lane 1 : Forebrain (Mouse) Tissue Lysate
Lane 2 : Spinal Cord (Mouse) Tissue Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes -
Western blot - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)This image is courtesy of Chris Anderson, Wellcome Trust Sanger Institute, United KingdomLanes 1-3 : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml
Lane 4 : nNOS antibody at 1/2500 dilution
Lanes 1 & 4 : mouse forebrain
Lane 2 : mouse forebrain with Mouse nNOS (neuronal) (phospho S847) peptide (ab16981) at 1 µg/ml
Lane 3 : mouse forebrain with corresponding unmodified nNOS (neuronal) peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 160 kDa
Observed band size: 160 kDa -
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)This image is courtesy of Sophie Pezet, CNRS, Paris, FranceImmunostaining using Rabbit polyclonal to nNOS (neuronal) (phospho S847) (ab16650) on rat brain tissue sections (30 micron free floating). ab16650 was used at a dilution of 1/3000 and incubated for 18 hours at RT (in PBS triton 0.3%). Secondary Antibody Goat anti-rabbit alexa Fluor 488 was used at a dilution of 1/1000. The image shows cytoplasmic staining of CNS neurons with ab16650 in naïve rats; the staining being observed in the soma and processes of these neurons. The staining was quenched by pre-incubation with peptide against phospho S847 (ab16981), but not by the control peptide (ab57047) indicating that ab16650 is specific for nNos phosphorylated at S847. Protocol: Rats were perfused-fixed with 4% paraformaldehyde. Tissues were post-fixed overnight in the same fixative and then cryoprotected in 20% sucrose overnight. Following embedding in OCT and freezing, tissues were cut and immunostained using the 'free floating’ technique.
Protocols
Datasheets and documents
References (12)
ab16650 has been referenced in 12 publications.
- Sharma NM et al. Exercise training augments neuronal nitric oxide synthase dimerization in the paraventricular nucleus of rats with chronic heart failure. Nitric Oxide 87:73-82 (2019). PubMed: 30878404
- Smith ACW et al. Accumbens nNOS Interneurons Regulate Cocaine Relapse. J Neurosci 37:742-756 (2017). PubMed: 28123012
- Rice FL et al. Anatomy and immunochemical characterization of the non-arterial peptidergic diffuse dural innervation of the rat and Rhesus monkey: Implications for functional regulation and treatment in migraine. Cephalalgia 37:1350-1372 (2017). PubMed: 27852962
- Choi SR et al. Neuronal NOS Activates Spinal NADPH Oxidase 2 Contributing to Central Sigma-1 Receptor-Induced Pain Hypersensitivity in Mice. Biol Pharm Bull 39:1922-1931 (2016). PubMed: 27601184
- Niu X et al. ß3-adrenoreceptor stimulation protects against myocardial infarction injury via eNOS and nNOS activation. PLoS One 9:e98713 (2014). WB ; Mouse . PubMed: 24911015
Images
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Western blot - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)All lanes : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml
Lane 1 : Forebrain (Mouse) Tissue Lysate
Lane 2 : Spinal Cord (Mouse) Tissue Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control)
Lane 4 : Forebrain (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide at 1 µg/ml
Lane 5 : Spinal Cord (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (phospho S847) peptide at 1 µg/ml
Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control) with Mouse nNOS (neuronal) (phospho S847) peptide at 1 µg/ml
Lane 7 : Forebrain (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (unmodified) peptide at 1 µg/ml
Lane 8 : Spinal Cord (Mouse) Tissue Lysate with Mouse nNOS (neuronal) (unmodified) peptide at 1 µg/ml
Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control) with Mouse nNOS (neuronal) (unmodified) peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutesThis blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab16650 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Western blot - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)All lanes : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml
Lane 1 : Forebrain (Mouse) Tissue Lysate
Lane 2 : Spinal Cord (Mouse) Tissue Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (negative control)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes
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Western blot - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) This image is courtesy of Chris Anderson, Wellcome Trust Sanger Institute, United KingdomLanes 1-3 : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml
Lane 4 : nNOS antibody at 1/2500 dilution
Lanes 1 & 4 : mouse forebrain
Lane 2 : mouse forebrain with Mouse nNOS (neuronal) (phospho S847) peptide (ab16981) at 1 µg/ml
Lane 3 : mouse forebrain with corresponding unmodified nNOS (neuronal) peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 160 kDa
Observed band size: 160 kDa
-
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) This image is courtesy of Sophie Pezet, CNRS, Paris, FranceImmunostaining using Rabbit polyclonal to nNOS (neuronal) (phospho S847) (ab16650) on rat brain tissue sections (30 micron free floating). ab16650 was used at a dilution of 1/3000 and incubated for 18 hours at RT (in PBS triton 0.3%). Secondary Antibody Goat anti-rabbit alexa Fluor 488 was used at a dilution of 1/1000. The image shows cytoplasmic staining of CNS neurons with ab16650 in naïve rats; the staining being observed in the soma and processes of these neurons. The staining was quenched by pre-incubation with peptide against phospho S847 (ab16981), but not by the control peptide (ab57047) indicating that ab16650 is specific for nNos phosphorylated at S847. Protocol: Rats were perfused-fixed with 4% paraformaldehyde. Tissues were post-fixed overnight in the same fixative and then cryoprotected in 20% sucrose overnight. Following embedding in OCT and freezing, tissues were cut and immunostained using the 'free floating’ technique.
