AB261889

Human CLDN1 knockout A-431 cell line

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CLDN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.7%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

5 Images

Lab

Western blot - Human CLDN1 knockout A-431 cell line (AB261889)

Lanes 1 - 4 : Merged signal (red and green). Green - ab211737 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab211737 was shown to specifically react with in wild-type A-431 cells as signal was lost in CLDN1 knockout A-431 cell line ab261889 (knockout cell lysate ab261698). Wild-type and A-431 CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab211737 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Claudin 1 antibody [EPRR18871] (<a href='/en-us/products/primary-antibodies/claudin-1-antibody-eprr18871-ab211737'>ab211737</a>) at 1/2000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CLDN1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CLDN1 knockout A-431 cell line (ab261889)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 22 kDa

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Lab

Western blot - Human CLDN1 knockout A-431 cell line (AB261889)

Lanes 1 - 3 : Merged signal (red and green). Green - ab180158 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab180158 was shown to specifically react with in wild-type A-431 cells as signal was lost in CLDN1 knockout A-431 cell line ab261889 (knockout cell lysate ab261698). Wild-type and A-431 CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab180158 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Claudin 1 antibody [EPR9306] (<a href='/en-us/products/primary-antibodies/claudin-1-antibody-epr9306-ab180158'>ab180158</a>) at 1/10000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CLDN1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CLDN1 knockout A-431 cell line (ab261889)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 22 kDa

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Lab

Western blot - Human CLDN1 knockout A-431 cell line (AB261889)

Lanes 1 - 2 : Merged signal (red and green). Green - ab129119 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab129119 was shown to specifically react with in wild-type A-431 cells as signal was lost in CLDN1 knockout cell line ab261889 (knockout cell lysate ab261698). Wild-type and CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab129119 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Claudin 1 antibody (<a href='/en-us/products/primary-antibodies/claudin-1-antibody-ab129119'>ab129119</a>) at 1 µg/mL

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CLDN1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CLDN1 knockout A-431 cell line (ab261889)

Predicted band size: 22 kDa

Observed band size: 18 kDa

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Lab

Next Generation Sequencing - Human CLDN1 knockout A-431 cell line (AB261889)

2 bp deletion (allele 1) and 5 bp deletion (allele 2) after Leu16 of the WT protein

Lab

Next Generation Sequencing - Human CLDN1 knockout A-431 cell line (AB261889)

X = 5 bp deletion, 2 bp deletion

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.7%

Disease

Epidermoid Carcinoma

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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>" }, "NGS": { "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?

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Properties and storage information

Gene name

CLDN1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing, Western blot

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.

Claudin 1 also known as CLDN1 is a protein that plays a fundamental role in the formation of tight junctions in the epithelial cells. This claudin protein has a molecular weight of approximately 22 kDa. You can find it expressed in various tissues including the skin liver kidney and lungs. Claudin 1 belongs to the claudin family which consists of transmembrane proteins involved in sealing the space between adjacent cells therefore ensuring tissue integrity.

Biological function summary

This claudin protein contributes to establishing and maintaining the barrier function that controls paracellular transport which is the movement of substances between cells. Claudin 1 participates as a critical component of the tight junction complex and it interacts with other proteins such as occludin and zonula occludens. These interactions contribute to the regulation and maintenance of cell polarity and signaling.

Pathways

Claudin 1 is involved in the epithelial cell signaling pathways that influence cellular proliferation and differentiation. One significant pathway is the Wnt signaling pathway where claudin 1 cooperates with other claudins and proteins like β-catenin to regulate gene transcription. Additionally it is implicated in the MAPK pathway which involves signaling cascades that affect cell growth division and response to external stress.

Claudin 1 shows a strong association with cancers particularly hepatocellular carcinoma and melanoma. In these conditions changes in claudin 1 expression levels can affect tumor progression and metastasis. It also associates with other proteins like E-cadherin impacting cell adhesion and epithelial-mesenchymal transition in cancerous cells. Moreover abnormalities in claudin 1 expression relate to skin disorders such as ichthyosis which is marked by a dysfunction in the skin barrier.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information CLDN1

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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'.

For licensing inquiries, please contact partnerships@abcam.com

Human CLDN1 knockout A-431 cell line

Western blot - Human CLDN1 knockout A-431 cell line (AB261889)

Western blot - Human CLDN1 knockout A-431 cell line (AB261889)

Western blot - Human CLDN1 knockout A-431 cell line (AB261889)

Next Generation Sequencing - Human CLDN1 knockout A-431 cell line (AB261889)

Next Generation Sequencing - Human CLDN1 knockout A-431 cell line (AB261889)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Disease

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Handling procedures

Initial handling guidelines

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Target data

Product promise