iNOS Mouse Nitric Oxide Synthase Cell Based Antagonist Assay, Panlabs
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Inducible nitric oxide synthase (iNOS) is a member of a growing family of enzymes that catalyze the conversion of L-arginine to citrulline and nitric oxide. Like other NOS isoforms, iNOS requires NADPH, FAD and terahydrobiopterin. From diabetes to hypertension, cancer to drug addiction, stroke to intestinal motility, memory and learning disorders to septic shock, inflammatory and autoimmune diseases, sunburn to anorexia, male impotence to tuberculosis, there is probably no pathological condition where nitric oxide does not play an important role.
Family:
Nitrogen Metabolism
Sub Family:
Nitric Oxide Synthase
Class:
L-Arginine Turnover
Protein Name:
iNOS
Uniprot Number:
P35228
Gene Name:
NOS2
Gene ID:
4843
Gene Aliases:
iNOS|NOS|HEP-NOS
Organism:
Mouse
Construct Details:
Endogenous
Assay Information
Assay Type:
Functional
Assay Sub Type:
Cell Based
Functional Mode:
Antagonist
Detection Method:
Spectrophotometry
Measured Response:
Absorbance
Testing Information
Procedure Summary:
RAW 264.7 cells (ATCC, TIB-71) are used. Test compound and/or vehicle is pre-incubated with the cells (5 x 10^5 /ml) in DMEM modified buffer for 20 minutes. The cells are then subjected to stimulation with LPS (0.1 µg/ml) and IFN-g (50 U/ml) for 24 hours at 37°C, 5 % CO2 incubator. After incubation, cell supernatant is mixed with Griess reagent and the optical density at 545 nm is measured. Test compound-induced inhibition of 0.1 µg/ml LPS and 50 U/ml IFN-g-induced increase of iNOS by 50 percent or more (≥50%) indicates antagonist activity.
Incubation:
1 d at 37°C
Control Inhibitor:
1400w
Criteria For Significance:
≥50% Inhibition of 0.1 ug/ml LPS and 50 U/ml IFN-gamma increase of iNOS response
Test Concentration / Dose:
Compounds are screened at 10, 3, 1, 0.3, 0.1, 0.03, 0.01 and 0.003 μM
Test Sample Requirements:
75 μl of 10 mM stock or 1 mg (pre-weighed) powder (assuming FW ≦500 and purity 100%)
Minimum Order Quantity:
16
Turnaround Time
Standard:
20 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
Panlabs
Testing Location:
Taiwan - Taipei
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