Human LEF1 knockout Jurkat cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
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] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-LEF1 antibody [EP2030Y] (<a href='/en-us/products/primary-antibodies/lef1-antibody-ep2030y-ab53293'>ab53293</a>) 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
Predicted band size: 44 kDa
Observed band size: 40 kDa
false
- WB
Lab
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] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-LEF1 antibody [EPR2029Y] (<a href='/en-us/products/primary-antibodies/lef1-antibody-epr2029y-ab137872'>ab137872</a>) 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
Predicted band size: 44 kDa
Observed band size: 40 kDa
false
- NGS
Supplier Data
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
- NGS
Supplier Data
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%
Complementary Products
Primary Antibodies
AB137872
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LEF1 antibody [EPR2029Y] (AB137872)](https://content.abcam.com/products/images/lef1-antibody-epr2029y-ab137872--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img418641.jpg)
- 5
- 3
Reactivity data
Product details
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.
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 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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com