Analysis Note

ControlMacrophages, liver lysates

Application

Immunohistochemistry:
1:5000 (ABC) dilution of a previous lot was used on tissue sections (rat liver).

Immunocytochemistry:
1:50,000 dilution of a previous lot was used on cultured cells.

Western blot:
1:5,000 (ECL). The antibody has been used successfully on rat liver. It is not recommended for use on rat brain extracts since unidentified bands may be present.

Optimal working dilutions must be determined by end user.

Research CategoryNeuroscience

Anti-Nitric Oxide Synthase II Antibody is an antibody against Nitric Oxide Synthase II for use in IC, IH & WB.

Research Sub CategoryOxidative Stress

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Nitric Oxide Synthases are the only enzymes know to simultaneously require five bound cofactors, FAD, FMN, haem, tetrahydrobiopterin (BH4) and Ca++-Calmodulin (CAM). In mammals, there are three distinct genes for NOS, NOS I is neuronal, NOS II is cytokine inducible, and NOS III is endothelial. The enzymes exist as homodimers, each monomer consisting of two major domains: Nterminal oxygenase domain, which belongs to the class of haem-thiolate proteins, and Cterminal reductase domain, which is homologous to NADPH:P450 reductase. The interdomain linker between the oxygenase and reductase domains contains a CaMbinding sequence. Functionally NOS is an enzyme that catalyzes the stepwise conversion of the amino acid L-arginine to nitric oxide and L-citrulline. NOS activity stimulates cyclic GMP, inducing smooth muscle relaxation by increasing intracellular cGMP, which in turn inhibits calcium entry into the cell, and decreases intracellular calcium concentrations. This also stimulates a cGMP-dependent protein kinase that activates myosin light chain phosphatase, the enzyme that dephosphorylates myosin light chains, which leads to smooth muscle relaxation.

Immunogen

Epitope: C-terminus

C-terminal 14 amino acid peptide from mouse macrophage nitric oxide synthase.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Rabbit polyclonal serum in buffer containing liquid 0.05% sodium azide.

Unpurified

Quality

Evaluated by Western Blot on Mouse liver lysates.

Western Blot Analysis:
1:1000 dilution of this antibody detected INOS on 10 µg of Mouse liver lysates.

Specificity

Reactivity to human iNOS is approximately 1%.

Inducible nitric oxide synthase (iNOS, NOS-II). No cross-reaction with rat nNOS and eNOS by Western blot and immunohistochemistry.

Storage and Stability

Stable for 6 months at -20°C in undiluted aliquots from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

131 kDa