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Product name
Anti-nNOS (neuronal) (phospho S847) antibody
See all nNOS (neuronal) products (21) ...
Description
Rabbit polyclonal to nNOS (neuronal) (phospho S847)
Tested applications
IHC-Fr, IHC-FoFr, WBmore details
Cross reactivity
Reacts with
Mouse, Rat, Apteronotus leptorhynchus
Predicted to work with
Rabbit, Human, Xenopus laevis, Zebrafish
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Mouse nNOS (neuronal), phosphorylated at S847.
(Peptide available as ab169 81.)
Positive control
This antibody gave a positive signal in mouse forebrain, mouse spinal cord and mouse cortex tissue lysates. HeLa whole cell lysate was used as a negative control. nNOS is widely expressed in the nervous system: cerebrum, olfactory bulb, hippocampus, midbrain, cerebellum, pons, medulla oblongata, and spinal cord. It is also found in skeletal muscle, spleen, heart, kidney, and liver.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Metabolism >> Types of disease >> Cancer
Metabolism >> Pathways and Processes >> Metabolism processes >> Hypoxia
Cancer >> Cancer Metabolism >> Response to hypoxia
Neuroscience >> Neurotransmission >> Nitric Oxide >> NOS
Applications
Show applications key
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.
IHC-Fr: Use at an assay dependent concentration.
IHC-FoFr: 1/3000.
WB: Use a concentration of 1 µg/ml. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).Can be blocked with nNOS (neuronal) peptide - phospho S847 (ab16981).
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.
Target information above from: UniProt accessionP29475
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- 2310005C01Rik antibody
- bNOS antibody
- Constitutive NOS antibody
see all
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
Anti-nNOS (neuronal) (phospho S847) antibody images:
Western blot - 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 : Mouse Cortex Tissue Lysate
Lane 3 : Spinal Cord (Mouse) Tissue Lysate
Lane 4 : Forebrain (Mouse) Tissue Lysate
with
Lane 5 : Mouse Cortex Tissue Lysate with
Lane 6 : Spinal Cord (Mouse) Tissue Lysate with
Lane 7 : Forebrain (Mouse) Tissue Lysate with nNOS (neuronal) peptide at 1 µg/ml
Lane 8 : Mouse Cortex Tissue Lysate with nNOS (neuronal) peptide at 1 µg/ml
Lane 9 : Spinal Cord (Mouse) Tissue Lysate with nNOS (neuronal) peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP)
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 : 30 seconds
Western blot - 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
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (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 - nNOS (neuronal) (phospho S847) antibody (ab16650)
Lanes 1 - 3 : Anti-nNOS (neuronal) (phospho S847) antibody (ab16650) at 1 µg/ml Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom
Lane 4 : nNOS antibody at 1/2500 dilution
Lane 1 : mouse forebrain
Lane 2 : mouse forebrain with
Lane 3 : mouse forebrain with corresponding unmodified nNOS (neuronal) peptide at 1 µg/ml
Lane 4 : mouse forebrain
Lysates/proteins at 20 µg per lane.
Predicted band size : 160 kDa
Observed band size : 160 kDa
Immunohistochemistry (PFA perfusion fixed frozen sections) - nNOS (neuronal) (phospho S847) antibody (ab16650)
Immunostaining 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.
Sophie Pezet, CNRS, Paris, France
Immunohistochemistry (Frozen sections) - Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)
ab16650 staining nNOS (neuronal) (phospho S847) in 16 µm thick sections of Apteronotus leptorhynchus by Immunohistochemistry (Frozen sections).
Tissue was fixed in 2% paraformaldehyde, permeabilized using 0.3% Triton X-100, blocked with 3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS for 1 hour at 24°C and then incubated with ab16650 at a 1/100 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/200 dilution.
Image courtesy of Dr Ruxandra Sirbulescu by Abreview.
References for Anti-nNOS (neuronal) (phospho S847) antibody (ab16650)
This product has been referenced in:
- Garzón J et al. RGSZ2 Binds to the Neural Nitric Oxide Synthase PDZ Domain to Regulate Mu-Opioid Receptor-Mediated Potentiation of the N-Methyl-D-Aspartate Receptor-Calmodulin-Dependent Protein Kinase II Pathway. Antioxid Redox Signal 15:873-87 (2011). WB; Mouse.Read more (PubMed: 21348811) »
- Lekontseva O et al. Role of neuronal nitric-oxide synthase in estrogen-induced relaxation in rat resistance arteries. J Pharmacol Exp Ther 339:367-75 (2011).Read more (PubMed: 21807885) »
See all 6 publications for this product
Publishing research using ab16650? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"