Catalog # NK6550

nNOS Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

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.

References
Mount, P.F. et al. (2007) J Mo.l Cell. Cardiol. 42(2):271.
Fisslthaler, B. et al. (2008) Circ Res. 102:1520.
Tyryshkin, A. et al. (2009) J Biol Chem. 285(1):784.
Loot, A. E. et al. (2009) J Exp Med. 206(13):2889.

Western blot analysis of nNOS expression in adult mouse brain (lanes 1 & 3) and rat GC cells (lanes 2 & 4). The blots were probed with mouse monoclonal anti-nNOS (C-terminal region) at 1:1000 (lanes 1 & 2) or rabbit polyclonal anti-nNOS at 1:250 (lanes 3 & 4).

Labeling of nNOS phosphorylation in rat PC12 cells. The cells were probed with mouse monoclonal (mAb) nNOS (NM4011), and rabbit polyclonal (pAb) nNOS (C-terminal region), nNOS (Tyr-895)/eNOS (Tyr-657), and nNOS (Tyr-1326)/iNOS (Tyr-1055). The antibodies were detected using secondary antibody conjugated to DyLight® 594.



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
NM4011 Stasko, S. et al. (2013) J Appl Physiol. 114(11):1629. WB: mouse diaphragm, normal, deficient
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
eNOS (Tyr-657)/nNOS (Tyr-895), phospho-specific Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms140
iNOS (Tyr-1055)[conserved site], phospho-specific Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms, Ck130
nNOS (C-terminal region) Mouse mAb 50 μlWB, E, ICC, IHCHu, Rt, Ms155
nNOS (C-terminal regiom) Rabbit pAb50 μlWB, E, ICC, IHCHu, Rt, Ms155
Anti-Mouse Ig:HRP Donkey pAb 100 μlWB, EMs
Anti-Rabbit Ig Light ChainSpecific:HRP Mouse mAb 100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The nNOS phospho-regulation kit can be used to examine phosphorylation of nNOS (Tyr-895)/eNOS (Tyr-657) and nNOS (Tyr-1326)/iNOS (Tyr-1055) conserved sites relative to the total expression of nNOS. The kit includes rabbit polyclonal and mouse monoclonal antibodies to detect total nNOS, and secondary reagents for rabbit polyclonal and mouse monoclonal antibody detection.