Immunofluorescence analysis of iNOS was performed using 70% confluent log phase A549 cells treated with a combination of TNF-α (5 ng/ml), IFNγ (50 U/ml) and IL-1β (50 U/ml) for 18 h. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with iNOS Rabbit Polyclonal Antibody (PA3030A) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the untreated cells with no signal. Panel f represents the no primary control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Sheep, Cat, Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C K(1131) K G S A L E E P K A T R L(1144) of mouse macrophage NOS.|
|Storage buffer||whole serum diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200|
|Western Blot (WB)||1:2,000|
* 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.
PA3-030A detects inducible nitric oxide synthase (iNOS) from human and mouse tissues. This antibody does not detect endothelial NOS (eNOS) or brain NOS (bNOS).
PA3-030A has been successfully used in Western blot and immunohistochemistry (paraffin and frozen) procedures. By Western blot, this antibody detects an ~130 kDa protein representing iNOS in induced RAW 264.7 cell extracts.
The PA3-030A immunogen is a synthetic peptide corresponding to residues C K(1131) K G S A L E E P K A T R L(1144) of mouse macrophage NOS.
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
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) and 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.
|Not Applicable||Not Cited||
Agrimonolide from Agrimonia pilosa suppresses inflammatory responses through down-regulation of COX-2/iNOS and inactivation of NF-¿B in lipopolysaccharide-stimulated macrophages.
PA3-030A was used in western blot to determine suppression of inflammatory responses through down-regulation of COX-2/iNOS and inactivation of NF-kappaB in lipopolysaccharide-stimulated macrophages by agrimonolide from agrimonia pilosa
|Chen L,Teng H,Fang T,Xiao J||Phytomedicine : international journal of phytotherapy and phytopharmacology (23:846)||2016|
Bubble CPAP Support after Discontinuation of Mechanical Ventilation Protects Rat Lungs with Ventilator-Induced Lung Injury.
PA3-030A was used in western blot to study bubble CPAP support after ceasing mechanical ventilation can protect the lungs of rats with ventilator-induced lung injury
|Wu CS,Chou HC,Huang LT,Lin YK,Chen CM||Respiration; international review of thoracic diseases (91:171)||2016|
Expression of nitric oxide synthase isoforms in the ovine fetal brain: alteration by hormonal and hemodynamic stimuli.
PA3-030A was used in western blot to investigate the expression of NOS isoforms in the ovine fetal brain and the effect of cerebral hypoperfusion and hormonal stimuli
|Wood CE,Giroux D||Journal of the Society for Gynecologic Investigation (13:329)||2006|
Evaluation of antioxidant activity and inhibitory effect on nitric oxide production of some common vegetables.
PA3-030A was used in western blot to investigate the antioxidant activity of some vegetables
|Bor JY,Chen HY,Yen GC||Journal of agricultural and food chemistry (54:1680)||2006|
Suppression effect of soy isoflavones on nitric oxide production in RAW 264.7 macrophages.
PA3-030A was used in western blot to study the effect of soy isoflavones on nitric oxide production in RAW 264.7 macrophages and its mechanism
|Sheu F,Lai HH,Yen GC||Journal of agricultural and food chemistry (49:1767)||2001|
Mechanisms of vascular instability in a transgenic mouse model of sickle cell disease.
PA3-030A was used in western blot to investigate the vasculature in sickle cell disease by means of a transgenic mouse model
|Nath KA,Shah V,Haggard JJ,Croatt AJ,Smith LA,Hebbel RP,Katusic ZS||American journal of physiology. Regulatory, integrative and comparative physiology (279:R1949)||2000|
Metal-induced modulation of nitric oxide production in vitro by murine macrophages: lead, nickel, and cobalt utilize different mechanisms.
PA3-030A was used in western blot to study the mechanisms for the effects of metals on nitric oxide production in murine macrophages
|Tian L,Lawrence DA||Toxicology and applied pharmacology (141:540)||1996|
Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase.
PA3-030A was used in western blot to investigate the characteristics of nitric oxide synthase in vivo
|Bredt DS,Hwang PM,Glatt CE,Lowenstein C,Reed RR,Snyder SH||Nature (351:714)||1991|
Ameliorative effects of pentoxifylline on NOS induced by diabetes in rat kidney.
PA3-030A was used in immunohistochemistry - paraffin section to analyze NOS induced by diabetes in rat kidney and ameliorative effects of pentoxifylline
|Sönmez MF,Dündar M||Renal failure (38:605)||2016|
Geraniol attenuates 2-acetylaminofluorene induced oxidative stress, inflammation and apoptosis in the liver of wistar rats.
PA3-030A was used in immunohistochemistry - paraffin section to analyze attenuation of 2-acetylaminofluorene induced by oxidative stress, apoptosis and inflammation in the liver of wistar rats by geraniol
|Hasan SK,Sultana S||Toxicology mechanisms and methods (25:559)||2015|
Intra-articular resveratrol injection prevents osteoarthritis progression in a mouse model by activating SIRT1 and thereby silencing HIF-2¿.
PA3-030A was used in immunohistochemistry - paraffin section to investigate the effect of resveratrol on osteoarthritis in mice
|Li W,Cai L,Zhang Y,Cui L,Shen G||Journal of orthopaedic research : official publication of the Orthopaedic Research Society (33:1061)||2015|
The effect of pomegranate fruit extract on testosterone-induced BPH in rats.
PA3-030A was used in immunohistochemistry - paraffin section to study the effect of Pomegranate Fruit Extract (PFE) in a testosterone-induced benign prostatic hyperplasia model
|Ammar AE,Esmat A,Hassona MD,Tadros MG,Abdel-Naim AB,Guns ES||The Prostate (75:679)||2015|
|Not Applicable||Not Cited||
Genistein improves 3-NPA-induced memory impairment in ovariectomized rats: impact of its antioxidant, anti-inflammatory and acetylcholinesterase modulatory properties.
PA3-030A was used in immunohistochemistry - paraffin section to characterize how 3-NPA-induced memory impairment in ovariectomized rats is improved by Genistein and its anti-inflammatory, antioxidant, and acetylcholinesterase modulatory properties
|Menze ET,Esmat A,Tadros MG,Abdel-Naim AB,Khalifa AE||PloS one (10:null)||2015|
Local pulmonary immune responses in domestic cats naturally infected with Cytauxzoon felis.
PA3-030A was used in immunohistochemistry - paraffin section to describe the immune responses in the lungs of cats naturally infected with C. felis
|Frontera-Acevedo K,Sakamoto K||Veterinary immunology and immunopathology (163:1)||2015|
Infusion of bone marrow mononuclear cells reduces lung fibrosis but not inflammation in the late stages of murine silicosis.
PA3-030A was used in immunohistochemistry - paraffin section to test if bone marrow mononuclear cells administered in the late stages of silica-induced damage would reduce the remodeling process
|Lopes-Pacheco M,Ventura TG,de Oliveira HD,Monção-Ribeiro LC,Gutfilen B,de Souza SA,Rocco PR,Borojevic R,Morales MM,Takiya CM||PloS one (9:null)||2014|
Histopathologic, biochemical and genotoxic investigations on chronic sodium nitrite toxicity in mice.
PA3-030A was used in immunohistochemistry - paraffin section to study the toxicity caused by the long term sodium nitrite (NaNO2) consumption in mice
|Özen H,Kamber U,Karaman M,Gül S,Ataki¿i E,Özcan K,Ataki¿i O||Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie (66:367)||2014|
Cyclooxygenase-2 and inducible nitric oxide synthase expression in thyroid neoplasms and their clinicopathological correlation.
PA3-030A was used in immunohistochemistry - paraffin section to investigate the effect of COX-2 and iNOS on tumor progression
|Kim KH,Kim SH,Kim SH,Back JH,Park MJ,Kim JM||Journal of Korean medical science (21:1064)||2006|
Nitric oxide synthase in diabetic rat testicular tissue and the effects of pentoxifylline therapy.
PA3-030A was used in immunohistochemistry and western blot to determine the effects of pentoxifylline therapy and nitric oxide synthase in diabetic rat testicular tissue
|Sönmez MF,K¿l¿ç E,Karabulut D,Çilenk K,Deligönül E,Dündar M||Systems biology in reproductive medicine (62:22)||2016|
Pretreatment with mineralocorticoid receptor blocker reduces intestinal injury induced by ischemia and reperfusion: involvement of inhibition of inflammatory response, oxidative stress, nuclear factor ¿B, and inducible nitric oxide synthase.
PA3-030A was used in immunohistochemistry to study the roles of oxidative stress, NFkB and iNOS in the mechanism by which pretreatment with a mineralocorticoid receptor blocker protects against ischemia-reperfusion-induced intestinal injury
|Ozacmak HS,Ozacmak VH,Barut F,Arasl¿ M,Ucan BH||The Journal of surgical research (191:350)||2014|
GSPE is superior to NAC in the prevention of contrast-induced nephropathy: might this superiority be related to caspase 1 and calpain 1?
PA3-030A was used in immunohistochemistry to study the potential role of calpain-1 in the greater protective effect against contrast-induced nephropathy of grape seed proanthocyanidin extract as compared to N-acetyl cysteine
|Ulusoy S,Ozkan G,Mungan S,Orem A,Yulug E,Alkanat M,Yucesan FB||Life sciences (103:101)||2014|
Intravenous glutamine administration reduces lung and distal organ injury in malnourished rats with sepsis.
PA3-030A was used in immunohistochemistry to study the protective effects against lung and distal organ damage of the intravenous administration of glutamine to protein-energy malnourished rats suffering sepsis
|de Oliveira GP,Silva JD,de Araújo CC,Prota LF,Abreu SC,Madeira C,Morales MM,Takiya CM,Diaz BL,Capelozzi VL,Panizzutti R,Pelosi P,Rocco PR||Shock (Augusta, Ga.) (41:222)||2014|
Perspective on rhabdomyolysis-induced acute kidney injury and new treatment options.
PA3-030A was used in immunohistochemistry to study the protective effects of a grape seed proanthocyanidin extract in a rat model of acute kidney injury induced by rhabdomyolysis
|Ulusoy S,Ozkan G,Alkanat M,Mungan S,Yulu¿ E,Orem A||American journal of nephrology (38:368)||2013|
Inducible nitric oxide synthase (iNOS) in gingival tissues of chronic periodontitis with and without diabetes: immunohistochemistry and RT-PCR study.
PA3-030A was used in immunohistochemistry to study the effects of type 2 diabetes on the gingival expression of iNOS in individuals with chronic peridontitis
|Shaker O,Ghallab NA,Hamdy E,Sayed S||Archives of oral biology (58:1397)||2013|
How does colistin-induced nephropathy develop and can it be treated?
PA3-030A was used in immunohistochemistry to identify the mechanism of colistin-induced nephropathy
|Ozkan G,Ulusoy S,Orem A,Alkanat M,Mungan S,Yulug E,Yucesan FB||Antimicrobial agents and chemotherapy (57:3463)||2013|
Mechanisms of the protective effects of curcumin against indomethacin-induced gastric ulcer in rats.
PA3-030A was used in immunohistochemistry to study the molecular mechanisms underlying the protective effects of curcumin in a rat gastric ulcer model
|Morsy MA,El-Moselhy MA||Pharmacology (91:267)||2013|
|Not Applicable||Not Cited||
Multimechanistic antifibrotic effect of biochanin a in rats: implications of proinflammatory and profibrogenic mediators.
PA3-030A was used in immunohistochemistry to assess the antifibrotic of biochanin A
|Breikaa RM,Algandaby MM,El-Demerdash E,Abdel-Naim AB||PloS one (8:null)||2013|
Prognostic value of arginase-II expression and regulatory T-cell infiltration in head and neck squamous cell carcinoma.
PA3-030A was used in immunohistochemistry to study the prognostic value of arginase-II expression and regulatory T-cell infiltration in head and neck squamous cell carcinoma
|Bron L,Jandus C,Andrejevic-Blant S,Speiser DE,Monnier P,Romero P,Rivals JP||International journal of cancer (132:E85)||2013|
Effect of valproic acid treatment on penile structure in prepubertal rats.
PA3-030A was used in immunohistochemistry to study the histological effects of valproic acid on the penis in prepubertal rats
|Kutlu O,Cansu A,Karagüzel E,Gürgen SG,Koç O,Gür M,Ozgür GK||Epilepsy research (99:306)||2012|
Inducible nitric oxide synthase inhibition attenuates physical stress-induced lung hyper-responsiveness and oxidative stress in animals with lung inflammation.
PA3-030A was used in immunohistochemistry to study the role of iNOS in physical stress-induced lung hyper-responsiveness in animals with lung inflammation
|Marques RH,Reis FG,Starling CM,Cabido C,de Almeida-Reis R,Dohlnikoff M,Prado CM,Leick EA,Martins MA,Tibério IF||Neuroimmunomodulation (19:158)||2012|
Immunohistochemical study of the angiogenetic network of VEGF, HIF1¿, VEGFR-2 and endothelial nitric oxide synthase (eNOS) in human breast cancer.
PA3-030A was used in immunohistochemistry to study the expression of VEGF, VEGFR-2, HIF1-alpha and eNOS in invasive ductal carcinoma
|Kafousi M,Vrekoussis T,Tsentelierou E,Pavlakis K,Navrozoglou I,Dousias V,Sanidas E,Tsiftsis D,Georgoulias V,Stathopoulos EN||Pathology oncology research : POR (18:33)||2012|
Role of nitric oxide in vascular permeability after combined burns and smoke inhalation injury.
PA3-030A was used in immunohistochemistry to study the regulation of the changes in microvascular permeability by NO with the existence of severe burn and/or smoke inhalation injury
|Soejima K,Traber LD,Schmalstieg FC,Hawkins H,Jodoin JM,Szabo C,Szabo E,Virag L,Salzman A,Traber DL,Varig L||American journal of respiratory and critical care medicine (163:745)||2001|
GnRH agonist-suppressed expression of nitric oxide synthases and generation of peroxynitrite in adenomyosis.
PA3-030A was used in immunohistochemistry to investigate the role of gonadotrophin-releasing hormone agonists (GnRHa) during nitric oxide synthase expression and peroxynitrite generation in adenomyosis
|Kamada Y,Nakatsuka M,Asagiri K,Noguchi S,Habara T,Takata M,Kudo T||Human reproduction (Oxford, England) (15:2512)||2000|
Inducible and endothelial nitric oxide synthase expression during development of transplant arteriosclerosis in rat aortic grafts.
PA3-030A was used in immunohistochemistry to detect the expression of iNOS and eNOS during development of transplant arteriosclerosis in rat aortic grafts
|Akyürek LM,Fellström BC,Yan ZQ,Hansson GK,Funa K,Larsson E||The American journal of pathology (149:1981)||1996|
Experimental allergic encephalomyelitis in the rat is inhibited by aminoguanidine, an inhibitor of nitric oxide synthase.
PA3-030A was used in immunohistochemistry to investigate the role of nitric oxide production in the pathogenesis of rat experimental allergic encephalomyelitis
|Zhao W,Tilton RG,Corbett JA,McDaniel ML,Misko TP,Williamson JR,Cross AH,Hickey WF||Journal of neuroimmunology (64:123)||1996|
Scutellarin regulates microglia-mediated TNC1 astrocytic reaction and astrogliosis in cerebral ischemia in the adult rats.
PA3-030A was used in immunocytochemistry and western blot to test if scutellarin affects reactive astrocytes in rats with middle cerebral artery occlusion
|Fang M,Yuan Y,Rangarajan P,Lu J,Wu Y,Wang H,Wu C,Ling EA||BMC neuroscience (16:null)||2015|
The cytotoxic, neurotoxic, apoptotic and antiproliferative activities of extracts of some marine algae on the MCF-7 cell line.
PA3-030A was used in immunocytochemistry to study the effect of cytotoxic, neurotoxic, apoptotic and antiproliferative activities of marine algae extracts on MCF-7 cell line
|Kurt O,Ozdal-Kurt F,Tu¿lu MI,Akçora CM||Biotechnic and histochemistry : official publication of the Biological Stain Commission (89:568)||2014|