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Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA752011

Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08] - BSA and Azide free

Application

  • WB

  • IP

  • FC

  • IHC-P

  • IHC-Fr

  • IF-Tissue

  • mIHC

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Predicted reactivity

  • Cynomolgus monkey

  • Pig

Predicted species support after-sales service
Conjugation

  • unconjugated

Overview

Product Name

Occludin Recombinant Rabbit Monoclonal Antibody [JJ091-08] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N-terminal human Occludin.

Species Reactivity

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

Predicted species support after-sales service

Validated Applications

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

Molecular Weight

Predicted band size: 59 kDa

Positive Control

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

Conjugation

unconjugated

Clone Number

JJ091-08

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • 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

Target

Function

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.

Background References

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).

Sequence Similarity

Belongs to the ELL/occludin family.

Tissue Specificity

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

Post-translational Modification

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.

Subcellular Location

Cell membrane, tight junction.

UNIPROT

SwissProt: Q16625 Human

SwissProt: Q61146 Mouse

SwissProt: Q6P6T5 Rat

Synonyms

BLCPMG antibody

FLJ08163 antibody

FLJ18079 antibody

FLJ77961 antibody

FLJ94056 antibody

MGC34277 antibody

Occludin antibody

Ocln antibody

OCLN_HUMAN antibody

Tight junction protein occludin antibody

Images

  • <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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</a>) at 1/2,000 dilution was used in <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.

    ☑ Knockdown (KD)

    Western blot analysis of Occludin on different lysates with Rabbit anti-Occludin antibody (HA752011) 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 (HA752011) 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.

  • Western blot analysis of Occludin on Mouse smooth muscle tissue lysate 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 /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 25 seconds; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 59.1 kDa<br />Observed band size: 59 kDa

    Western blot analysis of Occludin on Mouse smooth muscle tissue lysate with Rabbit anti-Occludin antibody (ET1701-76) at 1/1,000 dilution.


    Lysates/proteins at 10 µg/Lane.
    Exposure time: 25 seconds; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1701-76, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 59.1 kDa
    Observed band size: 59 kDa

  • Western blot analysis of Occludin on Caco-2 cell lysate 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 /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 120 seconds; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1701-76" style="font-weight: bold;text-decoration: underline;">ET1701-76</a>, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 59.1 kDa<br />Observed band size: 59 kDa

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


    Lysates/proteins at 10 µg/Lane.
    Exposure time: 120 seconds; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1701-76, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 59.1 kDa
    Observed band size: 59 kDa

  • 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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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 (HA752011) 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 (HA752011) 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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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 (HA752011) 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 (HA752011) 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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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 (HA752011) 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 (HA752011) 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/HA752011" style="font-weight: bold;text-decoration: underline;">HA752011</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 (HA752011, 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).

  • 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

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