Human E2F1 knockout HCT116 cell line
- Advanced Validation
- What is this?
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E2F1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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
Dmel\CG6376, Dmel_CG6376, E(Sev-CycE)3A, E(var)3-93E, E2-promoter binding facto, E2F transcription factor 1, E2F1_HUMAN, E2f-PA, E2f-PB, E2f-PC, E2f1 E2F transcription factor 1, Evar(3)164, KIAA4009, OTTHUMP00000030661, PBR3, PRB-binding protein E2F-1, RBAP-1, RBBP-3, RBP 3, Retinoblastoma-associated protein 1, Retinoblastoma-binding protein 3, Transcription factor E2F1, drosE2F1, l(3)07172, l(3)j3B1, l(3)j3C2, l(3)rM729, mKIAA4009
- WB
Lab
Western blot - Human E2F1 knockout HCT116 cell line (AB287380)
Western blot : Anti-E2F1 antibody [EPR26698-49] (ab314311) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab314311 was shown to bind specifically to E2F1. A band was observed at 47 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in E2F1 knockout cell line. To generate this image, wild-type and E2F1 knockout HCT 116 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 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-E2F1 antibody [EPR26698-49] (<a href='/en-us/products/primary-antibodies/e2f1-antibody-epr26698-49-ab314311'>ab314311</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
E2F1 knockout HCT 116 cell lysate at 20 µg
Lane 3:
Wild-type HEK-293 ab259780 cell lysate at 20 µg
Lane 4:
E2F1 knockout HEK-293 <a href='/en-us/products/cell-lines/human-e2f1-knockout-hek-293-cell-line-ab269502'>ab269502</a> cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- WB
Lab
Western blot - Human E2F1 knockout HCT116 cell line (AB287380)
Western blot : Anti-E2F1 antibody [KH95] (ab4070) staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab4070 was shown to bind specifically to E2F1. A band was observed at 47 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in E2F1 knockout cell line. To generate this image, wild-type and E2F1 knockout HCT 116 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 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-E2F1 antibody [KH95] (<a href='/en-us/products/primary-antibodies/e2f1-antibody-kh95-ab4070'>ab4070</a>) at 1/500 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
E2F1 knockout HCT 116 cell lysate at 20 µg
Lane 3:
Wild-type HEK-293 ab259780 cell lysate at 20 µg
Lane 4:
E2F1 knockout HEK-293 <a href='/en-us/products/cell-lines/human-e2f1-knockout-hek-293-cell-line-ab269502'>ab269502</a> cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human E2F1 knockout HCT116 cell line (AB287380)
91 bp deletion (allele 1) and 151 bp deletion (allele 2) after Pro121 of the WT protein
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 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.
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
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
McCoY5a + 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
E2F1 plays a significant role in cell growth regulation and apoptosis. E2F1 functions within a complex often forming heterodimers with DP proteins. This combination enhances its ability to bind DNA and activate transcription of genes necessary for DNA replication and cell cycle progression. E2F1 also participates in the control of apoptosis by regulating the expression of pro-apoptotic genes balancing cell proliferation and death.
Pathways
E2F1 is deeply embedded in cell cycle regulatory pathways and the p53 signaling pathway. It closely interacts with proteins like RB (Retinoblastoma protein) and p53. RB protein regulates E2F1 activity by controlling its release while p53 helps in mediating the cell's response to DNA damage potentially leading to either cell cycle arrest or apoptosis. These interactions highlight E2F1's role in maintaining cellular homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F1 antibody [KH95] (AB4070)](https://content.abcam.com/products/images/e2f1-antibody-kh95-ab4070--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img134267.jpg)
Primary Antibodies
AB51074
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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