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Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08]

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Catalog# ET1701-76

Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08]

Application

  • WB

  • IP

  • FC

  • IHC-P

  • IHC-Fr

  • IF-Tissue

  • mIHC

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Cynomolgus monkey

  • Pig

予測される種属に対するアフターサービスのサポート
Conjugation

  • unconjugated

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

概要

製品名

Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08]

抗体の種類

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within N-terminal human Occludin.

種属反応性

Human, Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

予測される種属に対するアフターサービスのサポート

検証された応用例

WB, IP, FC, IHC-P, IHC-Fr, IF-Tissue, mIHC

分子量

Predicted band size: 59 kDa

陽性対照

Human colon tissue, mouse colon tissue, rat colon tissue, MCF-7 cell lysate, Mouse smooth muscle tissue lysate.

結合

unconjugated

クローン番号

JJ091-08

RRID

AB_3070241

同源性数据

点击查看

製品の特徴

形態

Liquid

濃度

保存方法

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.

保存用バッファー

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

アイソタイプ

IgG

精製方法

Protein A affinity purified.

応用希釈度

  • WB

  • 1:1,000-1:2,000

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

  • IHC-P

  • 1:100-1:500

  • IHC-Fr

  • 1:500

  • IF-Tissue

  • 1:500

  • mIHC

  • 1:50

ターゲット

機能

Occludin is an integral membrane protein closely associated with the tight junctions of epithelial and endothelial cells. Occludin is a tetraspan integral membrane protein in epithelial and endothelial tight junction (TJ) structures that can contain two extracellular loops. The protein exists in a variety of phosphorylated forms. Phosphorylation is involved in regulating both the localization and the function of occludin. Expression of occludin is up-regulated by poly-unsaturated fatty acids, increasing transendothelial cell resistance and reducing cellular permeability to large molecules. The level of occludin varies greatly depending on tissue; in brain tissue, occludin is highly expressed at cell-cell contact sites. Non-neural tissues show lower expression and discontinuous distribution. Up-regulation of epithelial occludin may play a role in enhancing paracellular permeability and be related to the damage to the tight junction.

背景文献

1. Cao S et al. Hydrogen sulfide attenuates brain edema in early brain injury after subarachnoid hemorrhage in rats: Possible involvement of MMP-9 induced blood-brain barrier disruption and AQP4 expression. Neurosci Lett 621:88-97 (2016).

2. Zhang C et al. Catalpol downregulates vascular endothelial-cadherin expression and induces vascular hyperpermeability. Mol Med Rep 13:373-8 (2016).

配列相同性

Belongs to the ELL/occludin family.

組織特異性

Localized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis.

翻訳後修飾

Dephosphorylated by PTPRJ. The tyrosine phosphorylation on Tyr-398 and Tyr-402 reduces its ability to interact with TJP1. Phosphorylation at Ser-490 also attenuates the interaction with TJP1.

亜細胞局在

Cell membrane, tight junction.

UNIPROT #

SwissProt: Q16625 Human

SwissProt: Q61146 Mouse

SwissProt: Q6P6T5 Rat

別名

BLCPMG antibody

FLJ08163 antibody

FLJ18079 antibody

FLJ77961 antibody

FLJ94056 antibody

MGC34277 antibody

Occludin antibody

Ocln antibody

OCLN_HUMAN antibody

Tight junction protein occludin antibody

画像

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Occludin on different lysates with Rabbit anti-Occludin antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Occludin KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 59 kDa<br />Observed band size: 59 kDa<br /><br />Exposure time: 180 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/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/2,000 dilution was used in K1803 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.

    ☑ Knockdown (KD)

    Western blot analysis of Occludin on different lysates with Rabbit anti-Occludin antibody (ET1701-76) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Occludin KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 59 kDa
    Observed band size: 59 kDa

    Exposure time: 180 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 (ET1701-76) at 1/2,000 dilution was used in 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.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Colon<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1:500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: Colon

    Sample: Frozen section

    Antibody concentration: 1:500

    Antigen retrieval: Not required

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Occludin antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/100 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/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/100 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 tissue with Rabbit anti-Occludin antibody (ET1701-76) at 1/100 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 (ET1701-76) at 1/100 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 colon tissue with Rabbit anti-Occludin antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/500 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/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/500 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 colon tissue with Rabbit anti-Occludin antibody (ET1701-76) at 1/500 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 (ET1701-76) at 1/500 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 colon tissue with Rabbit anti-Occludin antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/100 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/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/100 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 colon tissue with Rabbit anti-Occludin antibody (ET1701-76) at 1/100 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 (ET1701-76) at 1/100 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.

  • Flow cytometric analysis of Caco-2 cells labeling Occludin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>, 1μg/mL) (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).

    Flow cytometric analysis of Caco-2 cells labeling Occludin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-76, 1μg/mL) (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 Occludin on different lysates with Rabbit anti-Occludin antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF-7 cell lysate (10 µg/Lane)<br />Lane 2: Mouse smooth muscle tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 59 kDa<br />Observed band size: 59 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% 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/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</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 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Occludin on different lysates with Rabbit anti-Occludin antibody (ET1701-76) at 1/1,000 dilution.

    Lane 1: MCF-7 cell lysate (10 µg/Lane)
    Lane 2: Mouse smooth muscle tissue lysate (20 µg/Lane)

    Predicted band size: 59 kDa
    Observed band size: 59 kDa

    Exposure time: 2 minutes;

    12% 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 (ET1701-76) 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 1:300,000 dilution was used for 1 hour at room temperature.

  • Application: IF-Tissue<br /><br />Species: Mouse<br /><br />Site: colon<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-Tissue

    Species: Mouse

    Site: colon

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

ご注意ください: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

論文での実績

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