Immunofluorescent analysis of uNOS (green) in SK-N-MC cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a uNOS monoclonal antibody (NOS-3F7-B11 B5) (Product # MA3-030 ) at a dilution of 1:20 and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.
|Tested species reactivity||Bovine, Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse|
|Host / Isotype||Mouse / IgM|
|Immunogen||Purified bovine bNOS.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:50-1:500|
* 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.
MA3-030 detects brain nitric oxide synthase (bNOS), inducible NOS (iNOS) and epithelial NOS (eNOS) in bovine, human, mouse and rat tissues.
MA3-030 has been successfully used in Western blot, immunofluorescence, immunoprecipitation, and immunohistochemical experiments. By Western blot, this antibody detects an ~130 kDa band representing iNOS in samples first induced with interferon (IFN) and lipopolysaccarides (LPS), an ~155 kDa band in tissues expressing bNOS and an ~140 kDa band in tissues expressing eNOS. Immunohistochemical staining of NOS with MA3-030 yields a pattern consistent with that seen in the literature and depends on the tissue being studied and the localization of the isoforms present.
The MA3-030 antigen is purified bovine bNOS.
Nitric oxide (NO) is an inorganic, gaseous free radical that carries a variety of messages between cells including vasorelaxation, neurotransmission and cytotoxicity. NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO. Two constitutive isoforms, brain/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 isoforms contain calmodulin, NADPH, FAD, and FMN binding domains. iNOS is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. Cytokines such as IFNg, TNF, IL-1, IL-2, and LPS cause an increase in iNOS mRNA, protein, and activity levels. 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. bNOS and eNOS share ~50% homology and their enzymatic activity depends on binding to the calcium/calmodulin complex. iNOS, however, is tightly bound to calmodulin and acts independently of calcium levels. Both constitutive isoforms respond immediately to increased levels of calcium to produce low levels of NO over a short time. In vascular endothelial cells NOS appears to modulate vasotension and platelet aggregation. bNOS is found in neurons, peripheral nerve cells, macula densa, and pancreatic islet cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Captopril and telmisartan treatments attenuate cadmium-induced testicular toxicity in rats.
MA3-030 was used in immunohistochemistry to study the mechanism by which captopril and telmisartan protect against testicular toxicity induced by cadmium exposure
|Fouad AA,Jresat I||Fundamental and clinical pharmacology (27:152)||2013|
Silibinin ameliorates arsenic induced nephrotoxicity by abrogation of oxidative stress, inflammation and apoptosis in rats.
MA3-030 was used in immunohistochemistry to study the mechanisms by which silibinin protects against nephrotoxicity induced by arsenic
|Prabu SM,Muthumani M||Molecular biology reports (39:11201)||2012|
Hepatoprotective effect of coenzyme Q10 in rats with acetaminophen toxicity.
MA3-030 was used in immunohistochemistry to investigate the protective effect of coenzyme Q10 against acetaminophen toxicity in rat liver
|Fouad AA,Jresat I||Environmental toxicology and pharmacology (33:158)||2012|
Protective effect of telmisartan against cadmium-induced nephrotoxicity in mice.
MA3-030 was used in immunohistochemistry to investigate the protective effect of telmisartan against cadmium nephrotoxicity
|Fouad AA,Jresat I||Life sciences (89:29)||2011|
Ameliorative effects of telmisartan in diabetic rats with indomethacin-induced gastric ulceration.
MA3-030 was used in immunohistochemistry to investigate the potential therapeutic application of telmisartan in diabetes treatment
|Fouad AA,Al-Sultan AI,Yacoubi MT,Gomaa W||European journal of pharmacology (637:162)||2010|
Coenzyme Q10 treatment ameliorates acute cisplatin nephrotoxicity in mice.
MA3-030 was used in immunohistochemistry to investigate the protective effect of Coenzyme Q10 against cisplatin nephrotoxicity in mice
|Fouad AA,Al-Sultan AI,Refaie SM,Yacoubi MT||Toxicology (274:49)||2010|
Distribution of NOS in normoxic vs. hypoxic rat lung: upregulation of NOS by chronic hypoxia.
MA3-030 was used in immunohistochemistry to investigate the changes of uNOS in the lungs of rats stimulated by chronic hypoxia
|Xue C,Rengasamy A,Le Cras TD,Koberna PA,Dailey GC,Johns RA||The American journal of physiology (267:L667)||1994|
Nitric oxide protects cardiac sarcolemmal membrane enzyme function and ion active transport against ischemia-induced inactivation.
MA3-030 was used in western blot to demonstrate the protective effect of nitric oxide on cardiac SL NOSs and sodium/potassium-ATPase against ischemia-induced inactivation.
|Xu KY,Kuppusamy SP,Wang JQ,Li H,Cui H,Dawson TM,Huang PL,Burnett AL,Kuppusamy P,Becker LC||The Journal of biological chemistry (278:41798)||2003|