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eNOS Recombinant Rabbit Monoclonal Antibody [PSH16-96] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751610

eNOS Recombinant Rabbit Monoclonal Antibody [PSH16-96] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

eNOS Recombinant Rabbit Monoclonal Antibody [PSH16-96] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human eNOS aa 950-1,203.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC, IP

Molecular Weight

Predicted band size: 133 kDa

Positive Control

EA.hy926 cell lysate, bEnd.3 cell lysate, Mouse placenta tissue lysate, Mouse kidney tissue lysate, Rat placenta tissue lysate, Rat kidney tissue lysate, EA.hy926, human colon carcinoma tissue, human placenta tissue, human kidney tissue, human spleen tissue, mouse spleen tissue, rat kidney tissue, rat spleen tissue, rat placenta tissue.

Conjugation

unconjugated

Clone Number

PSH16-96

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:50

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Endothelial NOS (eNOS), also known as nitric oxide synthase 3 (NOS3) or constitutive NOS (cNOS), is an enzyme that in humans is encoded by the NOS3 gene located in the 7q35-7q36 region of chromosome 7. This enzyme is one of three isoforms that synthesize nitric oxide (NO), a small gaseous and lipophilic molecule that participates in several biological processes. The other isoforms include neuronal nitric oxide synthase (nNOS), which is constitutively expressed in specific neurons of the brain and inducible nitric oxide synthase (iNOS), whose expression is typically induced in inflammatory diseases. eNOS is primarily responsible for the generation of NO in the vascular endothelium, a monolayer of flat cells lining the interior surface of blood vessels, at the interface between circulating blood in the lumen and the remainder of the vessel wall. NO produced by eNOS in the vascular endothelium plays crucial roles in regulating vascular tone, cellular proliferation, leukocyte adhesion, and platelet aggregation. Therefore, a functional eNOS is essential for a healthy cardiovascular system.

Background References

1. Yao L et al. Non-steroidal mineralocorticoid receptor antagonist finerenone ameliorates mitochondrial dysfunction via PI3K/Akt/eNOS signaling pathway in diabetic tubulopathy. Redox Biol. 2023 Dec

2. Teng Z et al. eNOS polymorphisms on male infertility: An updated systematic review and meta-analysis. Medicine (Baltimore). 2023 Jun

Subcellular Location

Cell membrane, Membrane, caveola, Cytoplasm, cytoskeleton, Golgi apparatus.

UNIPROT

SwissProt: P29474 Human

SwissProt: P70313 Mouse

SwissProt: Q62600 Rat

Synonyms

cNOS antibody

Constitutive NOS antibody

EC NOS antibody

EC-NOS antibody

ecNOS antibody

Endothelial nitric oxidase synthase antibody

Endothelial nitric oxide synthase antibody

Endothelial nitric oxide synthase 3 antibody

Endothelial NOS antibody

eNOS antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/5,000 dilution.<br /><br />Lane 1: EA.hy926 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (negative) (20 µg/Lane)<br />Lane 3: bEnd.3 cell lysate (20 µg/Lane)<br />Lane 4: RAW264.7 cell lysate (negative) (20 µg/Lane)<br />Lane 5: Mouse placenta tissue lysate (30 µg/Lane)<br />Lane 6: Mouse kidney tissue lysate (30 µg/Lane)<br />Lane 7: Rat placenta tissue lysate (30 µg/Lane)<br />Lane 8: Rat kidney tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 133 kDa<br />Observed band size: 133 kDa<br /><br />Exposure time: Lane 1-2: 10 seconds; Lane 3-8: 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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.

    ☑ Relative expression (RE)

    Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody (HA751610) at 1/5,000 dilution.

    Lane 1: EA.hy926 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (negative) (20 µg/Lane)
    Lane 3: bEnd.3 cell lysate (20 µg/Lane)
    Lane 4: RAW264.7 cell lysate (negative) (20 µg/Lane)
    Lane 5: Mouse placenta tissue lysate (30 µg/Lane)
    Lane 6: Mouse kidney tissue lysate (30 µg/Lane)
    Lane 7: Rat placenta tissue lysate (30 µg/Lane)
    Lane 8: Rat kidney tissue lysate (30 µg/Lane)

    Predicted band size: 133 kDa
    Observed band size: 133 kDa

    Exposure time: Lane 1-2: 10 seconds; Lane 3-8: 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 (HA751610) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of EA.hy926 (positive) and HeLa (negative) labeling eNOS with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of EA.hy926 (positive) and HeLa (negative) labeling eNOS with Rabbit anti-eNOS antibody (HA751610) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-eNOS antibody (HA751610) at 1/50 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 (HA601187, 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 colon carcinoma tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</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 colon carcinoma tissue with Rabbit anti-eNOS antibody (HA751610) 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 (HA751610) 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 placenta tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</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 placenta tissue with Rabbit anti-eNOS antibody (HA751610) 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 (HA751610) 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 kidney tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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 kidney tissue with Rabbit anti-eNOS antibody (HA751610) at 1/50 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 (HA751610) at 1/50 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-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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-eNOS antibody (HA751610) at 1/50 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 (HA751610) at 1/50 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 mouse spleen tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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 mouse spleen tissue with Rabbit anti-eNOS antibody (HA751610) at 1/50 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 (HA751610) at 1/50 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 rat kidney tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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 rat kidney tissue with Rabbit anti-eNOS antibody (HA751610) at 1/50 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 (HA751610) at 1/50 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 rat spleen tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/50 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 rat spleen tissue with Rabbit anti-eNOS antibody (HA751610) at 1/50 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 (HA751610) at 1/50 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 rat placenta tissue with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</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 rat placenta tissue with Rabbit anti-eNOS antibody (HA751610) 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 (HA751610) 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Flow cytometric analysis of HeLa (left, negative) and EA.hy926 (right, positive) cells labeling eNOS.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    ☑ Relative expression (RE)

    Flow cytometric analysis of HeLa (left, negative) and EA.hy926 (right, positive) cells labeling eNOS.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751610, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody (<a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/5,000 dilution.<br /><br />Lane 1: His-tagged eNOS recombinant protein<br />Lane 2: His-tagged nNOS recombinant protein<br /><br />Lysates/proteins at 20 ng/Lane.<br /><br />Exposure time: 2 seconds; 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/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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.

    Western blot analysis of eNOS on different lysates with Rabbit anti-eNOS antibody (HA751610) at 1/5,000 dilution.

    Lane 1: His-tagged eNOS recombinant protein
    Lane 2: His-tagged nNOS recombinant protein

    Lysates/proteins at 20 ng/Lane.

    Exposure time: 2 seconds; 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 (HA751610) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • eNOS was immunoprecipitated from 0.2 mg bEnd.3 cell lysate with <a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a> at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: bEnd.3 cell lysate (input)<br />Lane 2: <a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a> IP in bEnd.3 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751610" style="font-weight: bold;text-decoration: underline;">HA751610</a> in bEnd.3 cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 3 minutes; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    eNOS was immunoprecipitated from 0.2 mg bEnd.3 cell lysate with HA751610 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751610 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: bEnd.3 cell lysate (input)
    Lane 2: HA751610 IP in bEnd.3 cell lysate
    Lane 3: Rabbit IgG instead of HA751610 in bEnd.3 cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 3 minutes; ECL: K1801

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