EnCor Biotechnology

Rabbit Polyclonal Antibody to Nitric Oxide Synthase 1 (NOS1, neuronal NOS, nNOS) Cat# RPCA-NOS1

$300.00
Description

     The RPCA-NOS1 antibody was made against amino acids 1404-1434 of human NOS1 coupled to keyhole limpet hemocyanin (KLH)through a Cys residue. The peptide is CTYEVTNRLRSESIAFIEESKKDTDEVFSS and is 100% conserved between mouse and human, with one conservative substitution in rat, see "additional data". We document that the antibody recognizes NOS1 in samples from human. rodents and many other mammals and works well for western blotting ICC and IHC. Th antibody recognizes a subset of neurons throughout the brain. Mouse select image at left for larger view.

 Collections:
Amount: 100µL of 1mg/mL
Amount: 100µL of 1mg/mL
Immunofluorescent analysis of rat cortex section stained with rabbit pAb to NOS1, RPCA-NOS1, dilution 1:3,000 in red, and costained with chicken pAb to GFAP, CPCA-GFAP, dilution 1:5,000 in green. The blue is Hoechst staining of nuclear DNA. Following transcardial perfusion with 4% paraformaldehyde, the brain was post fixed for 24 hours, cut to 40μM, and free-floating sections were stained with above antibodies. The NOS1 antibody stains certain types of neurons, while GFAP antibody labels astrocytes and other glial cells.
Western blot analysis of different tissue lysates using rabbit pAb to NOS1, RPCA-NOS1, dilution 1:2,000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord, [6] cow brain, and [7] cow spinal cord lysate. A band at about 150kDa corresponds to NOS1 protein is clearly seen in brain but not spinal cord extracts. A ~25kDa band detected only in mouse tissues is of unknown origin. We noticed no difference in ICC or IHC staining between mouse and rat tissues so this reactivity is likely irrelevant.

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Immunogen: The C-terminal peptide of human NOS1 amino acids 1406-1434 with an N-terminal Cys added allowing coupling to activated keyhole limpet hemocyanin.
HGNC Name: NOS1
UniProt: P29475
Molecular Weight: 161kDa
Host: Rabbit
Species Cross-Reactivity: Human, Rat, Mouse
RRID: Pending
Format: Immunogen affinity purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3
Applications: WB, IF/ICC, IHC
Recommended Dilutions: WB: 1:10,000-20,000. IF/ICC 1:2,000-5,000
Storage: Stable at 4°C for one year, for longer term store at -20°C

There is a family of nitric oxide synthases (NOS) which generate nitric oxide (NO) from L-arginine producing citrulline as a byproduct. The 1998 Nobel Prize in Physiology and Medicine was awarded jointly to Robert F. Furchgott, Louis J. Ignarro and Ferid Murad for the discovery of NOS family enzymes. NOS1 is one of the three isoforms of nitric oxide synthase, the others being NOS2 (inducible NOS, iNOS) and NOS3 (endothelial NOS, eNOS). NOS1 is also known as neuronal nitric oxide synthase (nNOS) responsible for in the production of nitric oxide (NO) in the nervous system. NO is a signaling molecule that helps with the regulation of blood flow, neurotransmission, and immune responses. NOS1 is primarily expressed in neurons, particularly in the central nervous system (CNS). It is involved in processes like long-term potentiation (LTP), a mechanism related to learning and memory. Dysfunction of NOS1 has been implicated in various neurological conditions, including Parkinson's disease, Alzheimer's disease, and schizophrenia, as well as in neuropathic pain and stroke.

     Below is the C-terminal peptide of human, rat and mouse NOS1. The sequences are identical except for a conservative Thr>Ala subsitution in the rat.


Human  TYEVTNRLRSESIAFIEESKKDTDEVFSS
Mouse  TYEVTNRLRSESIAFIEESKKDTDEVFSS
Rat    TYEVTNRLRSESIAFIEESKKDADEVFSS
       **********************:******




Chromogenic Immunostaining of a 4% paraformaldehyde fixed paraffin embedded rat cerebellum section with rabbit pAb to nitric oxide synthetase 1, RPCA-NOS1, dilution 1:2,000, detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. In this image, RPCA-NOS1 labels the cytoplasm of a subset of neurons within the molecular layer. This antibody performs well in testing with both 4% PFA and standard NBF fixed rat, mouse and human tissues, but failed to stain overly fixed human brain samples. Mouse select image for larger view.



Confocal image stack of a region of rat cortex stained with RPCA-NOS1 diluted 1:2,000 in red and costained with chicken polyclonal antibody to GFAP CPCA-GFAP diluted 1:5,000 in green. The RPCA-NOS1 antibody shows the processes and perikarya of a subset of neurons while the GFAP antibody reveals astrocytic cells. Nuclei of neurons and non-neuronal cells are revealed with DAPI stain in blue. Mouse select image for larger view.

1. Hall A V et al. Structural organization of the human neuronal nitric oxide synthase gene (NOS1). J. Biol. Chem. 269:33082-33090 (1994).
2. Solanki K et al. The expanding roles of neuronal nitric oxide synthase (NOS1). Peer J.13651.

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