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Immunofluorescence analysis of nNOS was done on 70% confluent log phase SH-SY5Y cells. 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 nNOS Rabbit Polyclonal Antibody (PA3032A) at 2ug/ml 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 (Product # 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Human, Mouse, Rabbit, Rat|
|Published species reactivity||Rat, Mouse, Human, Guinea pig|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues T(724) K R R A I G F K K L A E A V K(739) C of rat bNOS.|
|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))||1:800|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200-400|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||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.
PA3-032A detects brain nitric oxide synthase (bNOS) from bovine, mouse, rabbit and rat tissues. This antibody does not detect endothelial NOS (eNOS) or inducible NOS (iNOS).
PA3-032A has been successfully used in Western blot, immunoprecipitation and immunohistochemical procedures. By Western blot, this antibody detects an ~155 kDa protein representing bNOS from rat brain extract. Immunohistochemical staining of bNOS in rat brain with PA3-032A results in staining of the anterior pituitary.
The PA3-032A immunogen is a synthetic peptide corresponding to residues T(724) K R R A I G F K K L A E A V K(739) C of rat nNOS.
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.
eNOS and bNOS share ~50% sequence homology, and their activity depends on binding to the calcium/calmodulin complex. Both constitutive isoforms respond immediately to increased levels of calcium to produce low levels of NO over a short period of time. bNOS has been found in neurons, peripheral nerve cells, macula densa & pancreatic islet cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Regulation of vascular tone in animals overexpressing the sarcolemmal calcium pump.
PA3-032A was used in western blot to investigate a new regulatory mechanism of vascular tone through sarcolemmal calcium pump PMCA.
|Schuh K,Quaschning T,Knauer S,Hu K,Kocak S,Roethlein N,Neyses L||The Journal of biological chemistry (278:41246)||2003|
Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase.
PA3-032A 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|
The plasmamembrane calmodulin-dependent calcium pump: a major regulator of nitric oxide synthase I.
PA3-032A was used in immunoprecipitation to identify the relationship between plasma membrane calcium/calmodulin-dependent calcium ATPase and nitric oxide synthase I.
|Schuh K,Uldrijan S,Telkamp M,Rothlein N,Neyses L||The Journal of cell biology (155:201)||2001|
Physiological dilation of uteroplacental arteries in the guinea pig depends on nitric oxide synthase activity of extravillous trophoblast.
PA3-032A was used in immunohistochemistry to investigate the role of nitric oxide synthase in trophoblast invasion in guinea pigs
|Nanaev A,Chwalisz K,Frank HG,Kohnen G,Hegele-Hartung C,Kaufmann P||Cell and tissue research (282:407)||1995|
bNOS; bNOS1; Brain NOS; constitutive NOS; N-NOS; NC-NOS; Neuronal Nitric Oxide Synthase; neuronal NOS; nitric oxidase synthase; nitric oxide synthase 1 (neuronal); nitric oxide synthase, brain; nNOS; NOS Type 1; NOS type I; peptidyl-cysteine S-nitrosylase NOS1
2310005C01Rik; Bnos; IHPS1; N-NOS; NC-NOS; nNOS; NO; NOS; Nos-1; NOS-I; NOS1