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iNOS Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER1706-89

iNOS Rabbit Polyclonal Antibody

Application

  • IF-Cell

  • FC

  • WB

  • IHC-Fr

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Chicken

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

iNOS Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within human iNOS aa 1,104-1,153 / 1,153.

Species Reactivity

Human, Mouse, Rat (Predicted: Chicken)

Predicted species support after-sales service

Validated Applications

IF-Cell, FC, WB, IHC-Fr

Molecular Weight

Predicted band size: 131 kDa

Positive Control

RAW264.7 treated with 1μg/mL LPS for 24 hours whole cell lysate, RAW264.7 whole cell lysate, HeLa cell lysate, A549 cell lysate, A549, LOVO.

Conjugation

unconjugated

RRID

AB_3069098

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IHC-Fr

  • 1:100

Target

Function

Nitric oxide (NO) has a broad range of biological activities and has been implicated in signaling pathways in phylogenetically diverse species. Nitric oxide synthases (NOSs), the enzymes responsible for synthesis of NO, contain an N-terminal oxygenase domain and a C-terminal reductase domain. NOS activity requires homodimerization as well as three cosubstrates (L-arginine, NADPH and O2) and five cofactors or prosthetic groups (FAD, FMN, calmodulin, tetrahydrobiopterin and heme). Several distinct NOS isoforms have been described and been shown to represent the products of three distinct genes. These include two constitutive Ca2+/CaM-dependent forms of NOS, including NOS1 (also designated ncNOS) whose activity was first identified in neurons, and NOS3 (also designated ecNOS), first identified in endothelial cells. The inducible form of NOS, NOS2 (also designated iNOS), is Ca2+-independent and is expressed in a broad range of cell types.

Background References

1. Hokari A et al. Cloning and functional expression of human inducible nitric oxide synthase (NOS) cDNA from a glioblastoma cell line A-172. J Biochem 116:575-581 (1994).

2. Guo F H et al. Continuous nitric oxide synthesis by inducible nitric oxide synthase in normal human airway epithelium in vivo. Proc Natl Acad Sci U.S.A. 92:7809-7813 (1995).

Sequence Similarity

Belongs to the NOS family.

Tissue Specificity

Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.

Post-translational Modification

Polyubiquitinated; mediated by SPSB1, SPSB2 and SPSB4, leading to proteasomal degradation.

Subcellular Location

Cytoskeleton, Nucleus, Cytosol.

UNIPROT

SwissProt: P35228 Human

SwissProt: P29477 Mouse

Synonyms

HEP-NOS antibody

Hepatocyte NOS antibody

HEPNOS antibody

inducible antibody

Inducible nitric oxide synthase antibody

Inducible NO synthase antibody

Inducible NOS antibody

iNOS antibody

MAC NOS antibody

Macrophage NOS antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of iNOS on different lysates with Rabbit anti-iNOS antibody (<a href="/products/ER1706-89" style="font-weight: bold;text-decoration: underline;">ER1706-89</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 treated with 1μg/mL LPS for 24 hours whole cell lysate (20 µg/Lane)<br />Lane 2: RAW264.7 whole cell lysate (20 µg/Lane)<br />Lane 3: HeLa cell lysate (30 µg/Lane)<br />Lane 4: A549 cell lysate (30 µg/Lane)<br /><br />Predicted band size: 131 kDa<br />Observed band size: 131 kDa<br /><br />Exposure time: 16 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER1706-89" style="font-weight: bold;text-decoration: underline;">ER1706-89</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of iNOS on different lysates with Rabbit anti-iNOS antibody (ER1706-89) at 1/1,000 dilution.

    Lane 1: RAW264.7 treated with 1μg/mL LPS for 24 hours whole cell lysate (20 µg/Lane)
    Lane 2: RAW264.7 whole cell lysate (20 µg/Lane)
    Lane 3: HeLa cell lysate (30 µg/Lane)
    Lane 4: A549 cell lysate (30 µg/Lane)

    Predicted band size: 131 kDa
    Observed band size: 131 kDa

    Exposure time: 16 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1706-89) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of RAW264.7 cells labeling iNOS with Rabbit anti-iNOS antibody (<a href="/products/ER1706-89" style="font-weight: bold;text-decoration: underline;">ER1706-89</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-iNOS antibody (<a href="/products/ER1706-89" style="font-weight: bold;text-decoration: underline;">ER1706-89</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of RAW264.7 cells labeling iNOS with Rabbit anti-iNOS antibody (ER1706-89) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-iNOS antibody (ER1706-89) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of C6 cells labeling iNOS with Rabbit anti-iNOS antibody (<a href="/products/ER1706-89" style="font-weight: bold;text-decoration: underline;">ER1706-89</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-iNOS antibody (<a href="/products/ER1706-89" style="font-weight: bold;text-decoration: underline;">ER1706-89</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling iNOS with Rabbit anti-iNOS antibody (ER1706-89) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-iNOS antibody (ER1706-89) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • ICC staining iNOS in LOVO cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining iNOS in LOVO cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: lung<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/100<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: lung

    Sample: Frozen section

    Antibody concentration: 1/100

    Antigen retrieval: Not required

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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