| 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. 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. Human iNOS is regulated by calcium/calmodulin (in contrast with mouse NOS2). |
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| Catalog # | N5350-10 |
| Applications | Suitable for use in Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications have not been tested. |
| Recommended Dilutions | Western Blot: 1-4ug/ml detects iNOS/NOS II (~125kD) in RIPA lysates from RAW 264.7 cells which had been stimulated with LPS. RAW 264.7 cell lysate that were treated with LPS, were resolved by electrophoresis, transferred to nitrocellulose and probed with N5350-10 (2ug/ml). Proteins were visualized using an IgG (HRP) goat anti-rabbit and a chemiluminescence detection system. |
| Immunohistochemistry: Reported to detect iNOS/NOS II in 2% paraformaldehyde-fixed sheep lung |
| Immunoprecipitation: 4ug immunoprecipitates iNOS/NOS II from 500ug of L929B cells which had been stimulated with 10ng/ml IFNg and 1ug/ml LPS. |
| Optimal dilutions to be determined by the researcher. |
| Storage and Stability | May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
| CAS Number | n/a |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Source | Mouse |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide. |
| Purity | Purified by Protein A affinity chromatography. |
| Immunogen | Partial fusion protein raised against the N-terminus of mouse iNOS/NOS II. |
| Specificity | Recognizes the N-terminus of mouse iNOS, Mr ~125kD. Does not crossreact with nNOS/ NOS I. Species Crossreactivity: bovine, sheep and guinea pig. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
