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AB264897

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

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(1 Publication)

TJP1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Western blot - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line (AB264897)
  • WB

Lab

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

All lanes:

Western blot - Anti-ZO1 tight junction protein antibody [BLR092G] (<a href='/en-us/products/primary-antibodies/zo1-tight-junction-protein-antibody-blr092g-ab276131'>ab276131</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 (ab264897) at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

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

Lab

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

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 ° 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 (ab264897)

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

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Sanger Sequencing - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line (AB264897)
  • Sanger seq

Unknown

Sanger Sequencing - Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line (AB264897)

Homozygous : 1 bp deletion in exon 2.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

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

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Complementary Products

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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Cell Lines & Lysates

AB257744

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

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

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Proteins & Peptides

AB114189

Recombinant Human Occludin protein

SDS-PAGE - Recombinant Human Occludin protein (AB114189)

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  • 0
Primary Antibodies

AB211737

Anti-Claudin 1 antibody [EPRR18871]

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-Claudin 1 antibody [EPRR18871] (AB211737)

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Reactivity data

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

We will provide viable cells that proliferate on revival.

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?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab264897-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab255448 Human wild-type HeLa cell line", "number":"AB264897-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab264897 Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line", "number":"AB264897-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab264897-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab264897 Human TJP1 (ZO1 tight junction protein) knockout HeLa cell line", "number":"AB264897-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
TJP1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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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Target data

See full target information TJP1
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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

International journal of colorectal disease 39:205 PubMed39702605

2024

AQP3 mediates autophagy through SIRT1/p62 signal to alleviate intestinal epithelial cell damage caused by sepsis.

Applications

Unspecified application

Species

Unspecified reactive species

Canmin Wang,Yingfang Hu,Yunfeng Song,Xinyi Hu
View all publications
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Product promise

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