Nitric oxide (NO), a diffusible free radical gas, 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.
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.
~130-160kD).
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/bNOS/NOS-I), endothelial (eNOS/NOS-III), and macrophage (mNOS/iNOS/NOS-II).
Both nNOS and eNOS are constitutive and the mNOS/iNOS is inducible.
Sequence homology among different cloned isoforms is ~ 50%.
Applications:Suitable for use in ELISA, Western Blotting.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000-1:5000 for neat serum and 1-10ug/ml for affinity pure using ECL.
A band of 95-110kD has been detected in brush border membrane (2)
ELISA:1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity purified.
Control peptide can be used to coat ELISA plates at 1ug/ml.
Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are stable for at least 12 months at -20°C.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
仕様
Size:100ul
Host:chicken
Source Antibody:rat
Grade:Serum
Purity:Serum
Form:Supplied as a liquid, neat serum, 40% glycerol.
Specificity:Recognizes NOS I, 17aa peptide sequence within the cytoplasmic, C-terminal domain of rat NOS I. Human, rat and mouse bNOS/NOS-1 are 1433aa, 1429aa and 1429aa proteins respectively. The rat NOS I peptide sequence is 95% conserved in mouse, human and rabbit. No significant sequence homology is seen with NOS-2/NOS-3 or other proteins.
Isotype:IgY
Calc Applications Abbrev:E WB
Calc Crossreactivity:Rt
Immunogen:A 17aa peptide within the cytoplasmic, C-terminal domain of the rat NOS I (1) conjugated to KLH.