Nitric oxide (NO), a diffusible free radical gas, affects a variety of physiological functions. NO acts as a neurotransmitter in brain and peripheral nervous system. It accounts for the activity of endothelium-derived relaxing factors, which stimulate vasodilatation by releasing NO from the endothelium. NO has been implicated in neurotoxicity associated with stroke and neurodegenerative diseases, neuronal regulation of smooth muscle including peristalsis, and penile erections. In macrophages, NO mediates tumoricidal and bactericidal actions. It mediates the actions of the excitatory neurotransmitter glutamate in stimulating cGMP levels. Unlike typical neurotransmitter, NO is a not stored in synaptic vesicle and does not act on membrane receptors. Synthesis of NO, initially demonstrated in vascular endothelium, is now found in many tissues.

NO is synthesized by L-arginine, oxygen, and NADPH by three known isoforms of heme-containing flavoproteins termed NO synthase (NOS, I-III, mol wt. ~130-160 kDa). One group of enzyme is constitutive, agonist-triggered, and dependent on Ca2+/Calmodulin and is inhibited by L-arginine analogues (L-NNA, L-NMMA). It is found in endothelium, adrenal glands, brain and platelets. The other principle group is inducible, Ca2+/Calmodulin-independent, and inhibited by NMMA and L-NNA. It has been found in macrophage, hepatocytes, tumor cells, vascular smooth muscle and endothelial cells. Analyses of cDNA clones have identified three distinct NOS genes in mammals: neuronal (nNOS), endothelial (eNOS), and macrophage (mNOS). Both nNOS and eNOS are constitutive and the mNOS/iNOS is inducible. Recently cloning of dNos (Drosophila NOS), an enzyme similar to constitutive NOSs, has been reported. Sequence homology among different cloned isoforms is ~ 50%.

ADI has produced antibodies to various NOS's using isoform specific peptide sequences. These antibodies do not crossreact with each other. Because of conservation of selected sequences between species (mouse, rat, human), antibodies show interspecies crossreactivity. The control peptides, used for immunization, are also available to determine specificity of antibodies. Appropriate cell/tissue extracts from natural cells/tissues and from recombinant baculovirus cells expressing high levels of a given NOS are available as well. These Extracts can be used as Positive controls for Western blotting.
 

M= Mouse; R=Rat; H=Human; Ha=Hamster; Rb=Rabbit; B=Bovine; C=Chicken; D=DOG; CT= near C-terminus; NT=near N-terminus; I=Middle of protein; CL=Cytoplasmic loop; EC=Extracellular domain; IC=Intracellular domain;

WB Positive controls- NOS have been expressed in baculovirus (Sf9) cells and purified (~95%) using column chromatography. Recombinant enzymes are supplied in 100-250 ul SDS-PAGE sample buffer (reduced). NOS preparations are not catalytically active and intended to be used as positive control for Western Blotting. ADI provides other preparations of NOSs that are enzymatically active.

cDNA Probes for various NOSs are also available (please inquire for details). cDNA probes may be labeled by radioactive or non-radioactive labels and suitable for use in Northern or Southern blotting.


"Neat Antisera" are the unpurified antiserum and it is suitable for ELISA and Western.
"Affinity pure" antibodies have been over the antigen-affinity column and recommended for immunohistochemical applications.
"Control peptides" can not be used for Western as they are very short peptides. They are intended for ELISA or antibody competition studies.

All Products are for in vitro research use only.