Human TLE1 knockout MCF7 cell line
- Advanced Validation
- What is this?
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TLE1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- ICC/IF
Unknown
Immunocytochemistry/ Immunofluorescence - Human TLE1 knockout MCF7 cell line (AB269498)
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 ab183742 at 1/500 dilution and 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) (ab150081) at 2 µg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (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™).
- WB
Lab
Western blot - Human TLE1 knockout MCF7 cell line (AB269498)
Lanes 1 - 4 : Merged signal (red and green). Green - ab183742 observed at 83 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
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 ab183742 and 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 (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-TLE 1 antibody [EPR9386(2)] (<a href='/en-us/products/primary-antibodies/tle-1-antibody-epr93862-ab183742'>ab183742</a>) 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
Predicted band size: 83 kDa
Observed band size: 83 kDa
false
- ICC/IF
Unknown
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 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) (ab150117) at 2 µg/ml (shown in green) and a goat secondary antibody to rabbit IgG (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).
- NGS
Supplier Data
Next Generation Sequencing - Human TLE1 knockout MCF7 cell line (AB269498)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%
- NGS
Supplier Data
Next Generation Sequencing - Human TLE1 knockout MCF7 cell line (AB269498)
1 bp insertion after Lys76 of the WT protein
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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.
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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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