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Immunofluorescent analysis of Phospho-nNOS pSer1417 (green) showing staining in the cytoplasm and nucleus of SH-SY5Y cells treated with 100 ng/ml nocodazole (left) and untreated SH-SY5Y cells (right). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-nNOS pSer1417 polyclonal antibody (Product # PA1-032) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Rabbit, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to residues C N(1411) R L R S E (pS) I A F I E E S K(1425) of human nNOS.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-032 detects phosphorylated nNOS (Ser 1417) from rat and human samples.
PA1-032 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~140 kDa protein representing recombinant rat phospho-nNOS (Ser 1417).
The PA1-032 immunogen is a synthetic phosphopeptide to residues C N(1411) R L R S E (pS) I A F I E E S K(1425) of human nNOS. The peptide sequence is 100% conserved in rat, frog, and mouse protein. This peptide (Cat. # PEP-188) is available for use in neutralization and control experiments.
Nitric oxide (NO) is an inorganic, gaseous free radical that carries a variety of messages between cells. Vasorelaxation, neurotransmission and cytotoxicity can all be potentiated through cellular response to NO. NO production is mediated by members of the nitric oxide synthase (NOS) family. NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO. Two constitutive isoforms, brain or neuronal NOS (b or nNOS, type I) & endothelial cell NOS (eNOS, type III), and one inducible isoform (iNOS, type II), have been cloned. All NOS isoforms contain calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN) binding domains.
iNOS is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. Cytokines such as interferon-gamma (IFN), tumor necrosis factor (TNF), interleukin-1 and -2, and lipopolysaccarides (LPS) cause an increase in iNOS mRNA, protein, and activity levels. Protein kinase C-stimulating agents exhibit the same effect on iNOS activity. After cytokine induction, iNOS exhibits a delayed activity response which is then followed by a significant increase in NO production over a long period of time.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Differential modulation by vascular nitric oxide synthases of the ethanol-evoked hypotension and autonomic dysfunction in female rats.
PA1-032 was used in western blot to study the role different NOS isoforms within the vasculature in hypotension and autonomic dysfunction induced by ethanol
|El-Mas MM,Fan M,Abdel-Rahman AA||Alcohol (Fayetteville, N.Y.) (46:727)||2012|
Enhancement of rostral ventrolateral medulla neuronal nitric-oxide synthase-nitric-oxide signaling mediates the central cannabinoid receptor 1-evoked pressor response in conscious rats.
PA1-032 was used in western blot to study the role of nNOS in central CB1 receptor-mediated pressor responses
|Ibrahim BM,Abdel-Rahman AA||The Journal of pharmacology and experimental therapeutics (341:579)||2012|
Sex differences in nitrosative stress during renal ischemia.
PA1-032 was used in western blot to investigate the differences between sexes in nitrosative stress during renal ischemia
|Rodríguez F,Nieto-Cerón S,Fenoy FJ,López B,Hernández I,Martinez RR,Soriano MJ,Salom MG||American journal of physiology. Regulatory, integrative and comparative physiology (299:R1387)||2010|
NO-dependent protective effect of VEGF against excitotoxicity on layer VI of the developing cerebral cortex.
PA1-032 was used in immunohistochemistry to study the mechanism for the neuroprotective effect of VEGF in the developing cerebral cortex
|El Ghazi F,Desfeux A,Brasse-Lagnel C,Roux C,Lesueur C,Mazur D,Remy-Jouet I,Richard V,Jégou S,Laudenbach V,Marret S,Bekri S,Prevot V,Gonzalez BJ||Neurobiology of disease (45:871)||2012|
constitutive NOS; N-NOS; NC-NOS; neuronal NOS; nitric oxidase synthase; nitric oxide synthase 1 (neuronal); nitric oxide synthase, brain; nNOS; NOS type I; peptidyl-cysteine S-nitrosylase NOS1; Phospho-bNOS; Phospho-Brain NOS; Phospho-Neural Nitric Oxide Synthase; Phospho-Nitric Oxide Synthase
Bnos; IHPS1; N-NOS; NC-NOS; nNOS; NOS; NOS1