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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

  • IF-Cell

  • IHC-P

  • IHC-Fr

  • IF-Tissue

  • mIHC

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Cynomolgus monkey

  • Pig

预测的种属支持售后
Conjugation

  • unconjugated

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

概述

产品名称

Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within N-terminal human Occludin.

种属反应性

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

预测的种属支持售后

验证应用

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

分子量

Predicted band size: 59 kDa

阳性对照

Human colon tissue, mouse colon tissue, rat colon tissue, Caco-2, MCF-7 cell lysate, Mouse smooth muscle tissue lysate.

偶联

unconjugated

克隆号

JJ091-08

RRID

AB_3070241

产品特性

形态

Liquid

存放说明

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.

存储缓冲液

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

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000-1:2,000

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

  • IF-Cell

  • 1:500

  • IHC-P

  • 1:100-1:500

  • IHC-Fr

  • 1:500

  • IF-Tissue

  • 1:500

  • mIHC

  • 1:50

靶点

功能

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).

序列相似性

Belongs to the ELL/occludin family.

组织特异性

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

翻译后修饰

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.

亚细胞定位

Cell membrane, tight junction.

UNIPROT

SwissProt: Q16625 Human

SwissProt: Q61146 Mouse

SwissProt: Q6P6T5 Rat

别名

BLCPMG antibody

FLJ08163 antibody

FLJ18079 antibody

FLJ77961 antibody

FLJ94056 antibody

MGC34277 antibody

Occludin antibody

Ocln antibody

OCLN_HUMAN antibody

Tight junction protein occludin antibody

图片

  • <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.

  • Immunocytochemistry analysis of Caco-2 cells labeling Occludin 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 />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-Occludin antibody (<a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>) at 1/500 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 Caco-2 cells labeling Occludin with Rabbit anti-Occludin antibody (ET1701-76) at 1/500 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-Occludin antibody (ET1701-76) at 1/500 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.

  • 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

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

引文

Filter:
  1. WB
  2. IF
  3. IHC-P
  4. IHC
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