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iNOS Recombinant Rabbit Monoclonal Antibody [PSH03-69]

Usd: 385

Specification

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Catalog# HA722031

iNOS Recombinant Rabbit Monoclonal Antibody [PSH03-69]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

iNOS Recombinant Rabbit Monoclonal Antibody [PSH03-69]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 131 kDa

Positive Control

RAW264.7 treated with 1μg/mL LPS for 24 hours cell lysate, RAW264.7 cells treated with 1μg/mL LPS for 6 hours, uman breast carcinoma tissue, human lung tissue, human spleen tissue.

Conjugation

unconjugated

Clone Number

PSH03-69

RRID

AB_3096842

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:200

  • IF-Cell

  • 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/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 treated with 1μg/mL LPS for 24 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 131 kDa<br />Observed band size: 131 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<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/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 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 (HA722031) at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 treated with 1μg/mL LPS for 24 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 3 minutes; ECL: K1801;

    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 (HA722031) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of RAW264.7 cells treated with or without 1μg/mL LPS for 6 hours labeling iNOS with Rabbit anti-iNOS antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% precooled methanol 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/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of RAW264.7 cells treated with or without 1μg/mL LPS for 6 hours labeling iNOS with Rabbit anti-iNOS antibody (HA722031) at 1/100 dilution.

    Cells were fixed in 100% precooled methanol 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 (HA722031) 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.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-iNOS antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-iNOS antibody (HA722031) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722031) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-iNOS antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-iNOS antibody (HA722031) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722031) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-iNOS antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722031" style="font-weight: bold;text-decoration: underline;">HA722031</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-iNOS antibody (HA722031) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722031) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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Citation