iNOS Antibody
iNOS Polyclonal Antibody for IF, IHC, Western blot
Synonyms:
Inducible Nitric Oxide Synthase, NOS Type II
Entrez Gene ID:
UniProt ID:
(Hu) P35228, (Ms) P29477, (Rt) Q06518
| Details | |
| Host / Isotype: | Rabbit |
| Class: | Polyclonal |
| Type: | Antibody |
| Species Reactivity: | Human (Hu) Mouse (Ms) Rat (Rt) |
| Immunogen: | Synthetic peptide corresponding to residues D(17) L K E E K D I N N N V K K T(31) of mouse iNOS. |
| Ordering Information | ||||
| Pierce iNOS Antibody |
| Storage: | -20° C, Avoid Freeze/Thaw Cycles |
| Form: | 200 µl of purified IgG (4.0 mg/ml) in PBS containing 1 mg/ml BSA and 0.05% sodium azide. |
| Applications | Dilution * |
| Immunofluorescence (IF) | 1:50 |
| Western Blot (WB) | 1:200 - 1:800 |
| Immunohistochemistry (IHC) | 1:20 |
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* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
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| Product Specific Information |
| PA1-036 detects inducible nitric oxide synthase (iNOS) from human, mouse and rat tissues and cells as well as recombinant human and mouse iNOS. This antibody does not detect other NOS isoforms. PA1-036 has been successfully use in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~135 kDa protein representing recombinant human iNOS and human iNOS from cytokine stimulated A549 cells. By Western blot PA1-036 also detects purified recombinant mouse iNOS, mouse iNOS from cytokine stimulated RAW 264.7 cells and cytokine stimulated rat fibroblast iNOS, though the signals are not as strong as those seen with the human samples. The PA1-036 immunogen is a synthetic peptide corresponding to residues D(17) L K E E K D I N N N V K K T(31) of mouse iNOS. |
| General Information |
| 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. |
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iNOS Antibody (PA1-036) in IF
Immunofluorescence with anti-iNOS Polyclonal Antibody (PA1-036)
Immunofluorescent analysis of iNOS in A549 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a iNOS polyclonal antibody (Product # PA1-036) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). iNOS staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification. -
iNOS Antibody (PA1-036) in IF
Immunofluorescence with anti-iNOS Polyclonal Antibody (PA1-036)
Immunofluorescent analysis of iNOS in NIH-3T3 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a iNOS polyclonal antibody (Product # PA1-036) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). iNOS staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification. -
iNOS Antibody (PA1-036) in IHC
Immunohistochemistry with anti-iNOS Polyclonal Antibody (PA1-036)
Immunohistochemistry was performed on normal deparaffinized human Heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing iNOS (Product #PA1-036) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting. -
iNOS Antibody (PA1-036) in IHC
Immunohistochemistry with anti-iNOS Polyclonal Antibody (PA1-036)
Immunohistochemistry was performed on normal deparaffinized human Lung tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing iNOS (Product #PA1-036) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
| PubMed References: |
| Immunocytochemistry | ||
| Species / Dilution | Summary | |
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Ms / 0 |
PA1-036 was used in immunocytochemistry to study the role of Stat3 mammary gland regression
J Pathol. 2012 May;227(1):106-17.
"Conditional deletion of Stat3 in mammary epithelium impairs the acute phase response and modulates immune cell numbers during post-lactational regression." Author(s): Hughes K, Wickenden JA, Allen JE, Watson CJ Number of Citations: 1 (See PubMed article  )
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| Immunohistochemistry | ||
| Species / Dilution | Summary | |
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Ms / Not Cited |
PA1-036 was used in immunohistochemistry to study the role of the activated macrophages during the T cell-mediated chronic psoriasiform skin inflammation
J Clin Invest. 2006 Aug;116(8):2105-14.
"Activated macrophages are essential in a murine model for T cell-mediated chronic psoriasiform skin inflammation." Author(s): Wang H, Peters T, Kess D, Sindrilaru A, Oreshkova T, Van Rooijen N, Stratis A, Renkl AC, Sunderkötter C, Wlaschek M, Haase I, Scharffetter-Kochanek K Number of Citations: 1 (See PubMed article )
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Ms / Not Cited |
PA1-036 was used in immunohistochemistry to investigate the effect of TNFalpha on fatty liver formation induced by fructose
J Nutr Biochem. 2011 Jun;22(6):527-34.
"Role of tumor necrosis factor ? (TNF?) in the onset of fructose-induced nonalcoholic fatty liver disease in mice." Author(s): Kanuri G, Spruss A, Wagnerberger S, Bischoff SC, Bergheim I Number of Citations: 5 (See PubMed article )
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Rt / 1:50 |
PA1-036 was used in immunohistochemistry and western blot to investigate the functional properties of iNOS in the rat AO and PA treated with endotoxin
Am J Physiol Regul Integr Comp Physiol. 2000 May;278(5):R1232-9.
"Differential inducible nitric oxide synthase expression in systemic and pulmonary vessels after endotoxin." Author(s): Pulido EJ, Shames BD, Fullerton DA, Sheridan BC, Selzman CH, Gamboni-Robertson F, Bensard DD, McIntyre RC Jr Number of Citations: 1 (See PubMed article )
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Rt / 1:500 |
PA1-036 was used in immunohistochemistry to investigate the effect of lung inflammation blockade on brain cooling after heatstroke injury
J Trauma. 2010 Oct;69(4):805-12.
"Inhibition of acute lung inflammation and injury is a target of brain cooling after heatstroke injury." Author(s): Hsi-Hsing Y, Ching-Ping C, Juei-Tang C, Lin MT Number of Citations: 0 (See PubMed article )
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| Western Blot | ||
| Species / Dilution | Summary | |
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Am / 1:1,000 |
PA1-036 was used in western blot to study the role of nitric oxide (NO) in the control of cardiac O2 consumption
Am J Physiol Heart Circ Physiol. 2004 Jul;287(1):H14-21.
"Endocardial endothelium in the avascular frog heart: role for diffusion of NO in control of cardiac O2 consumption." Author(s): Adler A, Huang H, Wang Z, Conetta J, Levee E, Zhang X, Hintze TH Number of Citations: 1 (See PubMed article )
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Hu / Not Cited |
PA1-036 was used in western blot to detect the expression of NOS2 in failing and nonfailing human hearts
Circulation. 2004 Apr 20;109(15):1886-91.
"Myocyte nitric oxide synthase 2 contributes to blunted beta-adrenergic response in failing human hearts by decreasing Ca2+ transients." Author(s): Ziolo MT, Maier LS, Piacentino V 3rd, Bossuyt J, Houser SR, Bers DM Number of Citations: 1 (See PubMed article )
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Hu / 1:4000 |
PA1-036 was used in western blot to study the role of reactive oxygen species in bile acid-induced hepatotoxicity
J Hepatol. 2011 Jul;55(1):133-44.
"Nitric oxide mimics transcriptional and post-translational regulation during ?-tocopherol cytoprotection against glycochenodeoxycholate-induced cell death in hepatocytes." Author(s): González R, Cruz A, Ferrín G, López-Cillero P, Fernández-Rodríguez R, Briceño J, Gómez MA, Rufián S, Mata Mde L, Martínez-Ruiz A, Marin JJ, Muntané J Number of Citations: 1 (See PubMed article )
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Ms / 1:200 |
PA1-036 was used in western blot to demonstrate that hypoxia is associated with increases in lung endothelial NOS in mice.
Respir Res. 2001;2(5):306-13.
"Upregulation of nitric oxide synthase in mice with severe hypoxia-induced pulmonary hypertension." Author(s): Fagan KA, Morrissey B, Fouty BW, Sato K, Harral JW, Morris KG Jr, Hoedt-Miller M, Vidmar S, McMurtry IF, Rodman DM Number of Citations: 5 (See PubMed article )
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Ms / Not Cited |
PA1-036 was used in western blot to study the role of class IA type PI3K in the bactericidal activity of macrophages.
J Biol Chem. 2006 Jun 30;281(26):17736-42.
"Phosphoinositide 3-kinase in nitric oxide synthesis in macrophage: critical dimerization of inducible nitric-oxide synthase." Author(s): Sakai K, Suzuki H, Oda H, Akaike T, Azuma Y, Murakami T, Sugi K, Ito T, Ichinose H, Koyasu S, Shirai M Number of Citations: 1 (See PubMed article )
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Ms / Not Cited |
PA1-036 was used in western blot to study the relationship between n-3 Polyunsaturated Fatty Acids and nitric oxide synthetase in hepatocytes
J Nutr Metab. 2011;2011():374542.
"Peroxidation of n-3 Polyunsaturated Fatty Acids Inhibits the Induction of iNOS Gene Expression in Proinflammatory Cytokine-Stimulated Hepatocytes." Author(s): Araki Y, Matsumiya M, Matsuura T, Oishi M, Kaibori M, Okumura T, Nishizawa M, Takada H, Kwon AH Number of Citations: 0 (See PubMed article )
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Rb / Not Cited |
PA1-036 was used in western blot to study the localization and function of the nitric oxide synthase in heart.
Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):657-62.
"Nitric oxide synthase in cardiac sarcoplasmic reticulum." Author(s): Xu KY, Huso DL, Dawson TM, Bredt DS, Becker LC Number of Citations: 35 (See PubMed article )
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Rt / 1:5,000 |
PA1-036 was used in western blot to investigate the characteristics of nitric oxide synthase in vivo
Nature. 1991 Jun 27;351(6329):714-8.
"Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase." Author(s): Bredt DS, Hwang PM, Glatt CE, Lowenstein C, Reed RR, Snyder SH Number of Citations: 1 (See PubMed article )
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Rt / 1:500 |
PA1-036 was used in immunohistochemistry and western blot to investigate the functional properties of iNOS in the rat AO and PA treated with endotoxin
Am J Physiol Regul Integr Comp Physiol. 2000 May;278(5):R1232-9.
"Differential inducible nitric oxide synthase expression in systemic and pulmonary vessels after endotoxin." Author(s): Pulido EJ, Shames BD, Fullerton DA, Sheridan BC, Selzman CH, Gamboni-Robertson F, Bensard DD, McIntyre RC Jr Number of Citations: 1 (See PubMed article )
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Rt / Not Cited |
PA1-036 was used in western blot to investigate the effect of sivelestat on iNOS expression in liver cells stimulated with interleukin 1beta
Dig Dis Sci. 2011 Jun;56(6):1672-81.
"Sivelestat suppresses iNOS gene expression in proinflammatory cytokine-stimulated hepatocytes." Author(s): Araki Y, Matsumiya M, Matsuura T, Kaibori M, Okumura T, Nishizawa M, Kwon AH Number of Citations: 0 (See PubMed article )
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(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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iNOS Antibodies > iNOS Polyclonal Antibody PA1-036
