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

iNOS Antibody in Immunohistochemistry (Paraffin) (IHC (P))

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iNOS Antibody — Detection of altered expression of the target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis using iNOS Recombinant Rat Monoclonal Antibody (CXNFT), Alexa Fluor™ Plus 488 Antibody (Product # 740025TP488), shows increased expression of iNOS upon treatment when compared to untreated cells. {TM}

Product Details

740025TP488

Applications

Tested Dilution

Publications

Western Blot (WB)

1:200

Immunohistochemistry (Paraffin) (IHC (P))

1:50

Immunocytochemistry (ICC/IF)

1:500

Flow Cytometry (Flow)

0.5 µg/test

Product Specifications

Species Reactivity

Mouse

Host/Isotype

Rat / IgG2a, kappa

Expression System

Expi293

Class

Recombinant Monoclonal

Type

Antibody

Clone

CXNFT

Conjugate

Alexa Fluor™ Plus 488 Alexa Fluor™ Plus 488 Alexa Fluor™ Plus 488

Excitation/Emission Max

493/518 nm View spectra

Form

Liquid

Concentration

1.0 mg/mL

50 µg

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

AB_3666230

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

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Gene Aliases: i-NOS; iNOS; Inosl; Nos-2; NOS-II; Nos2; Nos2a

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UniProt ID: (Mouse) P29477

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Entrez Gene ID: (Mouse) 18126

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Function(s)

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

Process(es)

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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iNOS Recombinant Rat Monoclonal Antibody (CXNFT), Alexa Fluor™ Plus 488

iNOS Antibody (740025TP488) in ICC/IF

Product Details

740025TP488

Western Blot (WB)

Immunohistochemistry (Paraffin) (IHC (P))

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Species Reactivity

Host/Isotype

Expression System

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Amount

Purification

Storage buffer

Contains

Storage conditions

Shipping conditions

RRID

Product Specific Information

Target Information

References (0)

Bioinformatics

Performance Guarantee

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