Catalog # NK6450

iNOS Phospho-Regulation Antibody Kit

Antibody Sampler Kit



Nitric oxide (NO) has a broad range of biological activities and is implicated in many cell signaling pathways. Nitric oxide synthases (NOS), the enzymes responsible for synthesis of NO, are fused enzymes that include a cytochrome reductase and a cytochrome that requires three cosubstrates (L-arginine, NADPH, and oxygen) and five cofactors (FAD, FMN, calmodulin, tetrahydrobiopterin, and heme). The NOS family includes two constitutive Ca2+/CaM-dependent forms of NOS: nNOS (also designated bNOS, NOS-I), whose activity was first identified in neurons and eNOS (also designated ecNOS, NOS-III) first identified in endothelial cells. The inducible form of NOS, iNOS (also designated NOS-II), is Ca2+ independent and is induced by microbial products in various cell types. Phosphorylation of NOS enzymes may regualte their activity. For example, Src kinase-induced phosphorylation of Tyr-1055 in iNOS reduces proteasomal degradation of iNOS, while shear stress induced phosphorylation of Tyr-657 in eNOS attenuates enzyme activity. Both of these tyrosine sites are conserved in nNOS, and may have important roles for regulate neuronal NOS activity.

Xie, Q.W. et al. (1992) Science 256:225.
Kleinert, H. et al. (2003) Biol Chem. 384(10-11):1343.
Musicki, B. et al. (2005) Proc. Natl. Acad. Sci.102(33):11870.
Tyryshkin, A. et al. (2009) J Biol Chem. 285(1):784.

Western blot analysis of mouse macrophages untreated (lanes 1 & 3) or treated with LPS (1µg/ml) for 18 hrs (lanes 2 & 4). The blots were probed with mouse monoclonal anti-iNOS at 1:500 (lanes 1 & 2) or rabbit polyclonal anti-iNOS at 1:250 (NP2131).

Western blot analysis of mouse macrophages (J774A.1) treated with LPS (1μg/ml) for 18 hrs followed by pervanadate (1 mM) for 30 min. (lanes 1, 3, & 5). The blots were then treated with alkaline phosphatase (lanes 2, 4, & 6). Blots were probed with rabbit polyclonal anti-inducible Nitric Oxide Synthase (iNOS) (lanes 1 & 2), anti-iNOS (Tyr-1055) (lanes 3 & 4), or mouse monoclonal anti-iNOS (lanes 5 & 6).

Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

iNOS(C-terminal region) Mouse mAb50 μlWB, E, ICCHu, Rt, Ms130
iNOS (C-terminal region) Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms130
iNOS (Tyr-1055)[conserved site], phospho-specific Rabbit pAb 50 μlWB, E, ICCHu, Rt, Ms130
Anti-Mouse Ig:HRPDonkey pAb100 μlWB, EMs
Anti-Rabbit Ig Light ChainSpecific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb

The inducible nitric oxide synthase (iNOS) phospho-regulation antibody sampler kit can be used to examine Tyr-1055 phosphorylation in iNOS. The kit also includes a mouse monoclonal and rabbit polyclonal antibody to monitor total expression levels of iNOS, as well as HRP-conjugated secondary reagents.