LEF1 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 = 4 bp insertion, 10 bp insertion, 4 bp deletion; Frameshift: 100%.
Images
Key facts
- Cell type
- Jurkat
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 4 bp insertion, 10 bp insertion, 4 bp deletion; Frameshift: 100%
Alternative names
DKFZp586H0919, FLJ46390, LEF1_HUMAN, Lymphoid enhancer-binding factor 1, T cell-specific transcription factor 1-alpha, TCF1-alpha, TCF10, TCF7L3, Transcription factor T cell specific 1 alpha
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LEF1 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 = 4 bp insertion, 10 bp insertion, 4 bp deletion; Frameshift: 100%.
Key facts
- Cell type
- Jurkat
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 4 bp insertion, 10 bp insertion, 4 bp deletion; Frameshift: 100%
- Disease
- Non-Hodgkin Lymphoma
- Concentration
Properties
- Gene name
- LEF1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Suspension
- Gender
- Male
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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 2x105 cells/mL is recommended.
- Do not allow cell density to exceed 3x106 cells/mL.
- Culture medium
- RPMI + 10% FBS
- 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
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
LEF1 also known as lymphoid enhancer-binding factor 1 is a transcription factor important in the Wnt signaling pathway. It typically weighs around 44 kDa. This protein contains a high-mobility group (HMG) box domain that enables it to bind specifically to DNA and influence gene expression. LEF1 is primarily expressed in tissues with active Wnt signaling including developing organs and certain cell types like T-cells specifically Jurkat cells. Researchers use LEF1 protein immunohistochemistry (LEF1 IHC) and LEF1 markers for expression analysis in different tissues.
- Biological function summary
The LEF1 transcription factor plays a significant role in cellular processes such as cell differentiation proliferation and embryonic development. As part of the Wnt signaling complex it interacts with β-catenin to regulate target gene expression. Activation of Wnt signaling by LEF1 can lead to changes in the cellular structure and function necessary for tissue development and regeneration. LEF1 also contributes to maintaining the pluripotency of stem cells by modulating gene networks critical for stem cell properties.
- Pathways
The LEF1 protein is essential in Wnt/β-catenin signaling and influences the Hedgehog pathway. LEF1 pairs with β-catenin to drive the transcription of Wnt target genes involved in cell cycle and differentiation. These pathways maintain stem cell populations and regulate development. LEF1's function in these pathways links it closely to other factors like TCF7L2 and GLI family proteins which also play vital roles within these signaling cascades.
- Associated diseases and disorders
LEF1 can be linked to conditions such as cancer and immune-related diseases. Overexpression or mutation of the LEF1 protein can lead to various types of cancers including leukemia and colorectal cancer by promoting abnormal cell proliferation and survival. LEF1's role in the immune system also associates it with autoimmune diseases where dysregulation of Wnt signaling might contribute to disease progression. Altered interactions between LEF1 and β-catenin or other associated proteins can disrupt normal cellular activities leading to these pathologies.
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4 product images
Western blot - Human LEF1 knockout Jurkat cell line (ab274898)
False colour image of Western blot: Anti-LEF1 antibody [EP2030Y] staining at 1/5000 dilution shown in black; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-LEF1 antibody [EP2030Y] ab53293 was shown to bind specifically to LEF1. A band was observed at 40/53 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lef1 knockout cell line ab274898 (knockout cell lysate ab274956). To generate this image wild-type and Lef1 knockout Jurkat cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times before development with Optiblot (ECL reagent Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged with 8 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-LEF1 antibody [EP2030Y] (Anti-LEF1 antibody [EP2030Y] ab53293) at 1/5000 dilution
Lane 1: Wild-type Jurkat cell lysate at 40 µg
Lane 2: Lef1 knockout Jurkat cell lysate at 40 µg
Lane 2: Western blot - Human LEF1 knockout Jurkat cell line (ab274898)
Lane 3: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 40 kDa
Western blot - Human LEF1 knockout Jurkat cell line (ab274898)
False colour image of Western blot: Anti-LEF1 antibody [EPR2029Y] staining at 1/1000 dilution shown in black; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-LEF1 antibody [EPR2029Y] ab137872 was shown to bind specifically to LEF1. A band was observed at 40/53 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lef1 knockout cell line ab274898 (knockout cell lysate ab274956). To generate this image wild-type and Lef1 knockout Jurkat cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times before development with Optiblot (ECL reagent Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged with 4 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (Anti-LEF1 antibody [EPR2029Y] ab137872) at 1/1000 dilution
Lane 1: Wild-type Jurkat cell lysate at 40 µg
Lane 2: Lef1 knockout Jurkat cell lysate at 40 µg
Lane 2: Western blot - Human LEF1 knockout Jurkat cell line (ab274898)
Lane 3: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 40 kDa
Next Generation Sequencing - Human LEF1 knockout Jurkat cell line (ab274898)
Knockout achieved by CRISPR/Cas9; X = 4 bp insertion, 10 bp insertion, 4 bp deletion; Frameshift: 100%
Next Generation Sequencing - Human LEF1 knockout Jurkat cell line (ab274898)
4 bp deletion after Pro133 (allele 1) and 4 bp deletion after Ile134 (allele 2) of the WT protein
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