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iNOS Antibody (740025TP488) in ICC/IF
Immunofluorescent analysis of iNOS was performed using 70% confluent untreated and LPS (1 µg/mL for 16h) treated RAW 264.7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were stained with iNOS Recombinant Rat Monoclonal Antibody (CXNFT), Alexa Fluor™ Plus 488 Antibody (Product # 740025TP488, 1:500) at 4 degree celsius overnight. Panel a) shows representative images of cells that were stained for detec... View More
Product Details
740025TP488
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2a, kappa
Expression System
Expi293
Class
Recombinant Monoclonal
Type
Antibody
Clone
CXNFT
Conjugate
Excitation/Emission Max
493/518 nm
View spectra
Form
Liquid
Concentration
1.0 mg/mL
Purification
Affinity chromatography
Storage buffer
proprietary buffer, pH 6.8
Contains
0.008% Bromonitrodioxane, 0.008% Methylisothiazolone
Storage conditions
4° C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
Product Specific Information
Alexa Fluor™ Plus recombinant antibodies are conjugated using new, proprietary dye chemistry so you can generate stunning data. Alexa Fluor™ Plus antibodies represent an advancement in fluorescent conjugate technology. Alexa Fluor™ Plus antibodies provide brighter signal compared to leading Alexa Fluor™ antibodies, providing you with better signal-to-noise for your critical experiments. These antibodies show better specificity and lot-to-lot consistency as these are recombinant antibodies, generated by cloning specific genes for the desired antibodies into an expression vector and expressed in vitro.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining.
Applications Tested: This CXNFT antibody has been tested by immunohistochemistry of mouse liver tissue and western blot, immunocytochemistry and flow cytometric analysis of RAW 264.7 cells treated with LPS. This may be used for immunocytochemistry at 2 µg/mL and for flow cytometry at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Excitation: 494 nm; Emission: 519 nm; Laser: Blue Laser
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
iNOS (Inducible Nitric oxide, NO, NOS) is an inorganic, gaseous free radical that carries a variety of messages between cells. Vasorelaxation, neurotransmission and cytotoxicity can all be potentiated through cellular response to NO. NO production is mediated by members of the nitric oxide synthase (NOS) family. iNOS is expressed in liver and inducible by a combination of lipopolysaccharide and certain cytokines. NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO. Two constitutive isoforms, brain or neuronal NOS (b or nNOS, type I) and endothelial cell NOS (eNOS, type III), and one inducible isoform (iNOS, type II), have been cloned. All NOS isoforms contain calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN) binding domains. iNOS is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. Cytokines such as interferon-gamma (IFN), tumor necrosis factor (TNF), interleukin-1 and -2, and lipopolysaccarides (LPS) cause an increase in iNOS mRNA, protein, and activity levels. Protein kinase C-stimulating agents exhibit the same effect on iNOS activity. After cytokine induction, iNOS exhibits a delayed activity response which is then followed by a significant increase in NO production over a long period of time. Three related iNOS pseudogenes are located within the Smith-Magenis syndrome region on chromosome 17. Diseases associated with iNOS dysfunction include achalasia and impotence.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: hepatocytes; inducible nitric oxide synthase; Inducible NO synthase; Inducible NOS; MAC-NOS; Macrophage NOS; nitric oxide synthase 2, inducible, macrophage; Nitric oxide synthase, inducible; nitric oxide synthase-inducible; NOS type II; OTTMUSP00000000202; Peptidyl-cysteine S-nitrosylase NOS2
Gene Aliases: i-NOS; iNOS; Inosl; Nos-2; NOS-II; Nos2; Nos2a
UniProt ID: (Mouse) P29477
Entrez Gene ID: (Mouse) 18126
actin binding
NADPH-hemoprotein reductase activity
nitric-oxide synthase activity
receptor binding
iron ion binding
protein binding
calmodulin binding
beta-catenin binding
cAMP-dependent protein kinase regulator activity
FMN binding
oxidoreductase activity
protein kinase binding
heme binding
tetrahydrobiopterin binding
arginine binding
protein homodimerization activity
cadherin binding
metal ion binding
flavin adenine dinucleotide binding
nitric-oxide synthase binding
Hsp90 protein binding
oxidoreductase
ovulation from ovarian follicle
response to hypoxia
arginine catabolic process
superoxide metabolic process
nitric oxide biosynthetic process
inflammatory response
nitric oxide mediated signal transduction
negative regulation of gene expression
peptidyl-cysteine S-nitrosylation
positive regulation of guanylate cyclase activity
response to lipopolysaccharide
cellular response to drug
regulation of cell proliferation
negative regulation of protein catabolic process
defense response to bacterium
negative regulation of blood pressure
regulation of protein kinase activity
positive regulation of vasodilation
regulation of insulin secretion
positive regulation of killing of cells of other organism
oxidation-reduction process
cellular response to lipopolysaccharide
cellular response to interferon-gamma
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