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AB257744

Human TJP1 (ZO1 tight junction protein) knockout HeLa cell lysate

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TJP1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.
2 Images
Western blot - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell lysate (AB257744)
  • WB

Lab

Western blot - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell lysate (AB257744)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : TJP1 knockout HeLa cell lysate 20 μg
Lane 3 : COS7 cell lysate 10 μg
Lane 4 : U-2 OS cell lysate 20 μg

Lanes 1 - 4 : Merged signal (red and green). Green - ab216880 observed at 200 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab216880 was shown to react with ZO1 tight junction protein in wild-type HeLa cells in Western blot with loss of signal observed in TJP1 knockout cell line ab264897 (TJP1 knockout cell lysate ab257744). Wild-type HeLa and TJP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab216880 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-ZO1 tight junction protein antibody (<a href='/en-us/products/primary-antibodies/zo1-tight-junction-protein-antibody-ab216880'>ab216880</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TJP1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-tjp1-zo1-tight-junction-protein-knockout-hela-cell-line-ab264897'>ab264897</a>)

Lane 3:

COS7 cell lysate at 10 µg

Lane 4:

U-2 OS cell lysate at 20 µg

Predicted band size: 195 kDa

Observed band size: 200 kDa

false

Sanger Sequencing - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell lysate (AB257744)
  • Sanger seq

Unknown

Sanger Sequencing - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell lysate (AB257744)

Homozygous : 1 bp deletion in exon 2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2.

Disease

Adenocarcinoma

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Complementary Products

Cell Lines & Lysates

AB264897

Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line

Advanced Validation

Western blot - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line (AB264897)

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Primary Antibodies

AB221547

Anti-ZO1 tight junction protein antibody [EPR19945-296]

20ul selling size|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZO1 tight junction protein antibody [EPR19945-296] (AB221547)

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  • 6
Primary Antibodies

AB224314

Anti-TJP2/ZO2 antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TJP2/ZO2 antibody (AB224314)

  • 0
  • 0
Primary Antibodies

AB18203

Anti-N Cadherin antibody - Intercellular Junction Marker

BOND RX™ Validated|Lab Essentials

Western blot - Anti-N Cadherin antibody - Intercellular Junction Marker (AB18203)

  • 4
  • 48
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
TJP1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The ZO1 tight junction protein also known as TJP1 is a critical component of tight junctions in epithelial and endothelial cells. It is a 225 kDa protein that plays a mechanical role in stabilizing cell-cell junctions by linking the actin cytoskeleton to tight junction membrane proteins. ZO1 is expressed in various tissues including the liver kidney and brain where it is essential for maintaining cell polarity and permeability barriers. It acts as a scaffold facilitating the organization of other tight junction proteins.
Biological function summary

ZO1 tight junction protein maintains cellular barriers and regulates paracellular permeability. This protein forms part of the tight junction complex with other proteins like occludin and claudins contributing to cellular integrity. ZO1's presence at tight junctions helps control the passage of ions and molecules between cells therefore ensuring selective permeability while protecting tissue function. Its ability to interact with multiple proteins enables it to coordinate complex signaling mechanisms that are important for cell adhesion and communication.

Pathways

The tight junction protein ZO1 participates in important signaling pathways like the epithelial barrier formation pathway and the actin cytoskeleton regulation pathway. It interacts with proteins like ZO2 and ZO3 which are critical for enhancing barrier function and cytoskeletal dynamics. By participating in these pathways ZO1 helps mediate both the structural and signaling functions necessary for maintaining tissue homeostasis and responding to cellular stress or injury.

ZO1 protein plays a role in conditions such as inflammatory bowel disease (IBD) and cancer. In IBD the disruption of ZO1 leads to compromised intestinal barriers contributing to disease pathogenesis. The loss or mislocalization of ZO1 has been associated with tumorigenesis in cancers where its interaction with other junctional proteins like claudins could influence cancer cell invasiveness. Understanding ZO1's involvement in these disorders provides insights into potential therapeutic targets for improving epithelial barrier function.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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