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ZO1 Recombinant Rabbit Monoclonal Antibody - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751628

ZO1 Recombinant Rabbit Monoclonal Antibody - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

ZO1 Recombinant Rabbit Monoclonal Antibody - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human ZO1 aa 1,401-1,748.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IF-Cell, IHC-P

Target Molecular Weight

Predicted band size: 195 kDa

Positive Control

A431 cell lysate, 293T cell lysate, HeLa cell lysate, U-2 OS cell lysate, C2C12 cell lysate, COS-1 cell lysate, MCF7, human kidney tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

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

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200

Target

Function

Zonula occludens-1 ZO-1, also known as Tight junction protein-1 is a 220-kD peripheral membrane protein that is encoded by the TJP1 gene in humans. It belongs to the family of zonula occludens proteins (ZO-1, ZO-2, and ZO-3), which are tight junction-associated proteins and of which, ZO-1 is the first to be cloned. It was first isolated in 1986 by Stevenson and Goodenough using a monoclonal antibody raised in rodent liver to recognise a 225-kD polypeptide in whole liver homogenates and in tight junction-enriched membrane fractions. It has a role as a scaffold protein which cross-links and anchors Tight Junction (TJ) strand proteins, which are fibril-like structures within the lipid bilayer, to the actin cytoskeleton. This gene encodes a protein located on a cytoplasmic membrane surface of intercellular tight junctions. The encoded protein may be involved in signal transduction at cell–cell junctions. Two transcript variants encoding distinct isoforms have been identified for this gene.

Background References

1. Tsurudome Y et al. Decreased ZO1 expression causes loss of time-dependent tight junction function in the liver of ob/ob mice. Mol Biol Rep. 2022 Dec

2. Han F et al. GLTSCR1 coordinates alternative splicing and transcription elongation of ZO1 to regulate colorectal cancer progression. J Mol Cell Biol. 2022 Jun

Subcellular Location

Cell membrane, Cell junction, tight junction, gap junction, Cell projection, podosome.

UNIPROT

SwissProt: Q07157 Human

SwissProt: P39447 Mouse

Entrez Gene: 292994 Rat

Synonyms

Tight junction protein 1 antibody

Tight junction protein ZO-1 antibody

Tight junction protein ZO1 antibody

TJP1 antibody

zo-1 antibody

Zo1 antibody

ZO1_HUMAN antibody

Zona occludens 1 antibody

Zona occludens 1 protein antibody

Zona occludens protein 1 antibody

Expand

Images

  • Western blot analysis of ZO1 on different lysates with Rabbit anti-ZO1 antibody (<a href="/products/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate<br />Lane 2: 293T cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: U-2 OS cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 195 kDa<br />Observed band size: 250 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<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/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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.

    Western blot analysis of ZO1 on different lysates with Rabbit anti-ZO1 antibody (HA751628) at 1/1,000 dilution.

    Lane 1: A431 cell lysate
    Lane 2: 293T cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: U-2 OS cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 195 kDa
    Observed band size: 250 kDa

    Exposure time: 3 minutes; ECL: K1802;
    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 (HA751628) 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.

  • Immunocytochemistry analysis of MCF7 cells labeling ZO1 with Rabbit anti-ZO1 antibody (<a href="/products/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</a>) at 1/100 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-ZO1 antibody (<a href="/products/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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.

    Immunocytochemistry analysis of MCF7 cells labeling ZO1 with Rabbit anti-ZO1 antibody (HA751628) at 1/100 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-ZO1 antibody (HA751628) 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 kidney tissue with Rabbit anti-ZO1 antibody (<a href="/products/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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 kidney tissue with Rabbit anti-ZO1 antibody (HA751628) 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 (HA751628) 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 mouse kidney tissue with Rabbit anti-ZO1 antibody (<a href="/products/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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 mouse kidney tissue with Rabbit anti-ZO1 antibody (HA751628) 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 (HA751628) 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 kidney tissue with Rabbit anti-ZO1 antibody (<a href="/products/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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/HA751628" style="font-weight: bold;text-decoration: underline;">HA751628</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 kidney tissue with Rabbit anti-ZO1 antibody (HA751628) 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 (HA751628) 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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