Immunofluorescent analysis of nNOS (green) showing staining in the cytoplasm and membrane of U251 cells (right) compared to a negative control without primary antibody (left). 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 an nNOS polyclonal antibody (Product # PA1-033) in 3% BSA-PBS at a dilution of 1:100 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, Mouse, Rat|
|Published species reactivity||Rat, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C N(1411) R L R S E S 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 *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||1:100-1:1000|
* 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.
PA1-033 detects nNOS from human, rat and mouse samples.
PA1-033 has been successfully used in Western blot, ICC/IF and immunohistochemisty (paraffin) procedures. By Western blot, this antibody detects an ~160 kDa protein. Detects a few non-specific bands in Western blot analysis of rat and mouse brain lysates. This antibody is not recommended for MCF-7, PC-12, SK-N-C or U87-MG cells in Western blot applications.
The PA1-033 immunogen is a synthetic peptide to residues C N(1411) R L R S E S I A F I E E S K(1425) of human nNOS. The peptide sequence is 100% conserved in rat, mouse, frog, and rabbit nNos proteins. This peptide (Cat. # PEP-190) 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) 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||
Effects of Aloe Vera on Spinal Cord Ischemia-Reperfusion Injury of Rats.
PA1-033 was used in immunohistochemistry - paraffin section to research spinal cord ischemia-reperfusion injury of rats and the effects of aloe vera
|Yuksel Y,Guven M,Kaymaz B,Sehitoglu MH,Aras AB,Akman T,Tosun M,Cosar M||Journal of investigative surgery : the official journal of the Academy of Surgical Research (29:389)||2016|
RNA silencing targeting PIN (protein inhibitor of neuronal nitric oxide synthase) attenuates the development of hypertension in young spontaneously hypertensive rats.
PA1-033 was used in western blot to study the role of PIN in the development of hypertension in a spontaneously hypertensive rat model using siRNA-mediated knock down
|Wang SC,Lin KM,Chien SJ,Huang LT,Hsu CN,Tain YL||Journal of the American Society of Hypertension : JASH (8:5)||2014|
Renal nitric oxide synthase and antioxidant preservation in Cyp1a1-Ren-2 transgenic rats with inducible malignant hypertension.
PA1-033 was used in western blot to study the role of renal NOS and antioxidant capacity in the maintenace of renal function in rats with inducible malignant hypertension
|Cunningham MW,Sasser JM,West CA,Milani CJ,Baylis C,Mitchell KD||American journal of hypertension (26:1242)||2013|
Aminoguanidine attenuates hypertension, whereas 7-nitroindazole exacerbates kidney damage in spontaneously hypertensive rats: the role of nitric oxide.
PA1-033 was used in western blot to study the ability of the nNOS pathway to protect against renal damage in a rat model of hypertension
|Huang CF,Hsu CN,Chien SJ,Lin YJ,Huang LT,Tain YL||European journal of pharmacology (699:233)||2013|
Nebivolol does not protect against 5/6 ablation/infarction induced chronic kidney disease in rats - comparison with angiotensin II receptor blockade.
PA1-033 was used in western blot to study the effect of a beta adrenergic receptor antagonist in two models of chronic kidney disease
|Sasser JM,Moningka NC,Tsarova T,Baylis C||Life sciences (91:54)||2012|
Endotoxemia exacerbates kidney injury and increases asymmetric dimethylarginine in young bile duct-ligated rats.
PA1-033 was used in western blot to study the effects of LPS on kidney damage and plasma asymmetric dimethylarginine in a rat model of cirrhosis
|Huang LT,Hung JF,Chen CC,Hsieh CS,Yu HR,Hsu CN,Tain YL||Shock (Augusta, Ga.) (37:441)||2012|
Aliskiren prevents hypertension and reduces asymmetric dimethylarginine in young spontaneously hypertensive rats.
PA1-033 was used in western blot to investigate the therapeutic effect of aliskiren for high blood pressure treatment
|Tain YL,Hsu CN,Lin CY,Huang LT,Lau YT||European journal of pharmacology (670:561)||2011|
Melatonin prevents hypertension and increased asymmetric dimethylarginine in young spontaneous hypertensive rats.
PA1-033 was used in western blot to investigate the therapeutic benefit of melatonin in hypertensive rats
|Tain YL,Huang LT,Lin IC,Lau YT,Lin CY||Journal of pineal research (49:390)||2010|
Renal nitric oxide production in rat pregnancy: role of constitutive nitric oxide synthases.
PA1-033 was used in western blot to investigate the renal nitric oxide production in rat pregnancy and possible involvement of different nitric oxide synthases
|Smith CA,Santymire B,Erdely A,Venkat V,Losonczy G,Baylis C||American journal of physiology. Renal physiology (299:F830)||2010|