NOS2/iNOS ELISA Kit (Nitric Oxide Synthase, Inducible)

INTENDED USE

Human NOS2/iNOS ELISA kit can measure concentrations of nitric oxide synthase, inducible (NOS2/iNOS, HEP-NOS. inducible NO) present is plasma, serum, biological fluids or tissue homogenate samples.

INTRODUCTION

Nitric oxide synthase 2, more commonly known as inducible nitric oxide synthase (iNOS or NOS2), is an enzyme critical to immune defense through its production of nitric oxide (NO). Unlike neuronal and endothelial NOS which exhibit transient activity, immune stimuli trigger sustained NO production by iNOS. This enables high output NO fluxes that potently combat pathogens but also risk unintended damage, necessitating tight control over iNOS expression and activity.

In general, proinflammatory cytokines induce iNOS expression while bacterial components and interferons synergize to further enhance activation. Once expressed, it dimerizes and oxidizes L-arginine to yield NO until substrate depletion intervenes. By inhibiting pathogen respiration, DNA replication, and protein function, NO elicits direct antimicrobial effects and synergizes with reactive oxygen intermediates. Furthermore, NO promotes vasodilation and leukocyte recruitment to infected sites. However, aberrant iNOS activity also contributes to septic shock and chronic inflammation underlying diseases like arthritis and colitis. Hence appropriate induction followed by silencing of iNOS once threats are cleared is imperative.

Though NO is too short-lived to be a viable therapeutic, targeting mechanisms governing iNOS activation shows promise against inflammatory disorders. For example, plant flavonoids like quercetin inhibit induction by suppressing NF-kB-mediated cytokine activity. Other small molecules may directly disrupt iNOS-dimerization or inhibit its catalytic activity rather than expression. Moreover, chemical modification of the iNOS promoter allows exogenous control over its induction, enabling site-specific regulation using gene therapy vectors.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-NOS2/iNOS antibody.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this NOS2/iNOS ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 78.125, 156.25, 312.5, 625, 1250, 2500, 5000 pg/ml.
Reactivity: Human
Sensitivity: 46.875 pg/ml
Range: 78.125 – 5000 pg/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for NOS2/iNOS, no cross reactivity or interference between NOS2/iNOS and analogues was detected.
– Recovery: Serum (85 – 102%), EDTA Plasma (89 – 103%), Heparin Plasma (88 – 102%).
– Linearity: Serum (86 – 105%), EDTA Plasma (84 – 94%), Heparin Plasma (81 – 100%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. LACC1 bridges NOS2 and polyamine metabolism in inflammatory macrophages. Nature. (2022) 609 (7926): 348-353. Wei Z., et al.
2. Nitric oxide and macrophage function. Annu Rev Immunol. (1997) 15: 323-50. MacMicking J., et al.
3. iNOS (NOS2) at a glance. J Cell Sci. (2004) 117 (Pt 14): 2865-7. Lowenstein C.J. and Padalko E.
4. Nitric-oxide synthase and neurodegeneration/neuroprotection. Proc Natl Acad Sci U S A. (1999) 96 (17): 9455-6. Kaufman P.L.

ADDITIONAL INFORMATION

• Full Name: NOS2/iNOS ELISA Kit (Nitric Oxide Synthase, Inducible)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 46.875 pg/ml

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