CLDN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.7%.
CLD1_HUMAN, Claudin-1, ILVASC, SEMP 1, Senescence associated epithelial membrane protein 1, Senescence-associated epithelial membrane protein
CLDN1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.7%.
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.
Recommended control: Human wild-type A-431 cell line (ab263975). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
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.
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.
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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Lanes 1 - 4: Merged signal (red and green). Green - Anti-Claudin 1 antibody [EPRR18871] ab211737 observed at 18 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-Claudin 1 antibody [EPRR18871] 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 Human CLDN1 knockout A-431 cell lysate ab261698). Wild-type and A-431 CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Anti-Claudin 1 antibody [EPRR18871] ab211737 and Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Claudin 1 antibody [EPRR18871] (Anti-Claudin 1 antibody [EPRR18871] ab211737) 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
Lanes 1 - 3: Merged signal (red and green). Green - Anti-Claudin 1 antibody [EPR9306] ab180158 observed at 18 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-Claudin 1 antibody [EPR9306] 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 Human CLDN1 knockout A-431 cell lysate ab261698). Wild-type and A-431 CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Anti-Claudin 1 antibody [EPR9306] ab180158 and Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Claudin 1 antibody [EPR9306] (Anti-Claudin 1 antibody [EPR9306] ab180158) 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
Performed under reducing conditions.
Predicted band size: 22 kDa
Lanes 1 - 2: Merged signal (red and green). Green - Anti-Claudin 1 antibody ab129119 observed at 18 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-Claudin 1 antibody 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 Human CLDN1 knockout A-431 cell lysate ab261698). Wild-type and CLDN1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Anti-Claudin 1 antibody ab129119 and Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Claudin 1 antibody (Anti-Claudin 1 antibody ab129119) 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
X = 5 bp deletion, 2 bp deletion
2 bp deletion (allele 1) and 5 bp deletion (allele 2) after Leu16 of the WT protein
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