iNOS Antibody [G10L10] | Primary Antibodies

Application: Reactivity: Mouse, Human, Rat

Usage Information

Dilution
WB1:1000 IP1:100 IF1:500

Application
WB, IP, IF, I-ELISA

Reactivity
Mouse, Human, Rat

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
131 kDa 131 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Mouse colon tissue; Mouse colon cancer tissue; RAW 264.7 cells (LPS, 0.1 µg/mL, 24 h ); L6 cells (IL-1β, 50 ng/ml; TNF-α, 20 ng/ml; IFN-γ, 100U/ml, 24 h); HepG2 cells (LPS, 10ug/ml, 6 h)
Negative Control	Mouse hippocampus tissue; RAW 264.7 cells; L6 cells; THP-1 cells

Datasheet & SDS

Biological Description

Specificity
iNOS Antibody [G10L10] detects endogenous levels of total iNOS protein.

Clone
G10L10

Synonym(s)
NOS2A; NOS2; Hepatocyte NOS; Inducible NO synthase; NOS type II; Peptidyl-cysteine S-nitrosylase NOS2; HEP-NOS; Inducible NOS; iNOS

Background
iNOS, or inducible nitric oxide synthase (NOS2), is the calcium-independent isoform of the NOS family. It forms a homodimeric enzyme of about 260 kDa per dimer that catalyzes the monooxygenation of L-arginine to produce nitric oxide (NO) and L-citrulline, using oxygen and NADPH. This reaction yields high-output NO, supporting sustained antimicrobial and inflammatory responses. Each 1153-residue monomer consists of three main regions: an N-terminal oxygenase domain (with heme b at His365, tetrahydrobiopterin at the active site, and a zinc tetrathiolate bridge via Cys1092 and Cys1098 that stabilizes the dimer), a calmodulin-binding hinge, and a C-terminal reductase domain (which contains FAD, FMN, and NADPH binding sites that enable electron transfer). Tight binding to calmodulin, even at low calcium concentrations (around 40 nM), locks iNOS in an active conformation without the need for further regulation. iNOS expression is transcriptionally induced by LPS and interferon gamma via the NF-kappaB, STAT1, and IRF1 pathways in macrophages, hepatocytes, and smooth muscle cells. The resulting enzyme generates cytotoxic and peroxynitrite-forming NO bursts that kill pathogens and signal through cGMP/PKG or S-nitrosylation. iNOS mediates innate immunity through M1 macrophage activation, supports vascular homeostasis, and promotes wound healing. Its activity can be inhibited by molecules like NAP110, which disrupt dimerization. Dysregulation of iNOS is implicated in septic shock (by inducing hypotension through guanylate cyclase activation), cancer (by promoting angiogenesis and suppressing T-cells), neurodegeneration (through nitration of amyloid beta and tau proteins), pain, and arthritis.

Background

iNOS, or inducible nitric oxide synthase (NOS2), is the calcium-independent isoform of the NOS family. It forms a homodimeric enzyme of about 260 kDa per dimer that catalyzes the monooxygenation of L-arginine to produce nitric oxide (NO) and L-citrulline, using oxygen and NADPH. This reaction yields high-output NO, supporting sustained antimicrobial and inflammatory responses. Each 1153-residue monomer consists of three main regions: an N-terminal oxygenase domain (with heme b at His365, tetrahydrobiopterin at the active site, and a zinc tetrathiolate bridge via Cys1092 and Cys1098 that stabilizes the dimer), a calmodulin-binding hinge, and a C-terminal reductase domain (which contains FAD, FMN, and NADPH binding sites that enable electron transfer). Tight binding to calmodulin, even at low calcium concentrations (around 40 nM), locks iNOS in an active conformation without the need for further regulation. iNOS expression is transcriptionally induced by LPS and interferon gamma via the NF-kappaB, STAT1, and IRF1 pathways in macrophages, hepatocytes, and smooth muscle cells. The resulting enzyme generates cytotoxic and peroxynitrite-forming NO bursts that kill pathogens and signal through cGMP/PKG or S-nitrosylation. iNOS mediates innate immunity through M1 macrophage activation, supports vascular homeostasis, and promotes wound healing. Its activity can be inhibited by molecules like NAP110, which disrupt dimerization. Dysregulation of iNOS is implicated in septic shock (by inducing hypotension through guanylate cyclase activation), cancer (by promoting angiogenesis and suppressing T-cells), neurodegeneration (through nitration of amyloid beta and tau proteins), pain, and arthritis.

References
https://pmc.ncbi.nlm.nih.gov/articles/PMC6908786/ https://pubmed.ncbi.nlm.nih.gov/21890489/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%