TLE1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 12 and 4 bp deletion in exon 12.
E(Sp1) homolog, ESG, ESG1, Enhancer of split groucho 1, Enhancer of split groucho-like protein 1, GRG1, TLE1_HUMAN, Transducin like enhancer of split 1, Transducin like enhancer of split 1 (E(sp1) homolog Drosophila), Transducin like enhancer of split 1 homolog of Drosophila E(sp1), Transducin-like enhancer protein 1
TLE1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 12 and 4 bp deletion in exon 12.
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.
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.
Transducin-Like Enhancer of Split 1 also known as TLE1 is a transcriptional corepressor well-known for its involvement in Notch and Wnt signaling pathways. TLE1 has a molecular mass of approximately 77 kDa. This protein is expressed in various tissues including the brain heart and lung highlighting its involvement in multiple biological systems. Researchers recognize its importance in modulating gene expression through interactions with different transcription factors.
The TLE1 protein plays a role in embryonic development by regulating the transcription of genes critical for cell differentiation. TLE1 forms part of a complex with other proteins to exert its corepressive function effectively. Through these interactions TLE1 is involved in differentiation processes in cells such as MCF-7 a breast cancer cell line influencing cellular behavior and function.
TLE1 influences the Notch and Wnt signaling pathways essential for cell fate determination and proliferation. Its interaction with key proteins like β-catenin and Notch receptor proteins connects it to these pathways modulating the transcription of target genes. TLE1's role in these pathways is important for maintaining balanced cellular processes and preventing abnormal cell growth.
TLE1 is linked to various forms of cancer including synovial sarcoma and breast cancer. Its overexpression has been observed in these cancers often in conjunction with other proteins like BCL2 supporting cell survival and resisting apoptosis. TLE1's involvement in these diseases makes it a target for therapeutic strategies aimed at modulating its activity to inhibit tumor progression.
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Anti-TLE 1 antibody [EPR9386(2)] ab183742 Anti-TLE 1 antibody [EPR9386(2)] was shown to specifically react with TLE 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264901 (knockout cell lysate Human TLE1 knockout HeLa cell lysate ab257241) was used. Wild-type and TLE 1 knockout samples were subjected to SDS-PAGE. Anti-TLE 1 antibody [EPR9386(2)] ab183742 and Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-TLE 1 antibody [EPR9386(2)] (Anti-TLE 1 antibody [EPR9386(2)] ab183742) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: TLE1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human TLE1 knockout HeLa cell line (ab264901)
Lane 3: 293T cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 83 kDa
Representative images of TLE1 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Allele-2: 1 bp deletion in exon 12.
Allele-1: 4 bp deletion in exon 12.
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