TLE1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%.
Images
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%
Alternative names
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
Recommended products
TLE1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%.
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- TLE1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- MEM + 10% FBS + 0.01 mg/ml bovine insulin
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Western blot data suggests that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type MCF7 cell line (ab271144). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Biological function summary
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.
- Pathways
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.
- Associated diseases and disorders
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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5 product images
Immunocytochemistry/ Immunofluorescence - Human TLE1 knockout MCF7 cell line (ab269498)
ab131648 staining TLE1 in wild-type MCF7 cells (top panel) and TLE1 knockout MCF7 cells (ab269498) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab131648 at 1/500 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor®488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 µg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor®594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 2 µg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Human TLE1 knockout MCF7 cell line (ab269498)
Anti-TLE 1 antibody [EPR9386(2)] ab183742 staining TLE1 in wild-type MCF7 cells (top panel) and TLE1 knockout MCF7 cells (ab269498) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-TLE 1 antibody [EPR9386(2)] ab183742 at 1/500 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 µg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 µg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).
Western blot - Human TLE1 knockout MCF7 cell line (ab269498)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-TLE 1 antibody [EPR9386(2)] ab183742 observed at 83 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.Anti-TLE 1 antibody [EPR9386(2)] ab183742 was shown to react with TLE 1 in western blot. The band observed in the CRISPR/Cas9 edited lysate lane below 83 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-TLE 1 antibody [EPR9386(2)] ab183742 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) 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 (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 in 20000 dilution for 1 h 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 MCF7 cell lysate at 20 µg
Lane 2: TLE1 CRISPR/Cas9 edited MCF7 cell lysate at 20 µg
Lane 2: Western blot - Human TLE1 knockout MCF7 cell line (ab269498)
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 83 kDa
Next Generation Sequencing - Human TLE1 knockout MCF7 cell line (ab269498)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%
Next Generation Sequencing - Human TLE1 knockout MCF7 cell line (ab269498)
1 bp insertion after Lys76 of the WT protein
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