Human E2F3 knockout HeLa cell line
- Advanced Validation
- What is this?
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E2F3 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 38 bp deletion in exon 2.
View Alternative Names
DKFZp686C18211, E2F transcription factor 3, E2F3_HUMAN, KIAA0075, MGC104598, Transcription factor E2F 3
- WB
Lab
Western blot - Human E2F3 knockout HeLa cell line (AB265362)
False colour image of Western blot : Anti-E2F3 antibody staining at 1/500 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab152126 was shown to bind specifically to E2F3. A band was observed at 49 kDa in wild-type HeLa cell lysates with no signal observed at this size in E2F3 knockout cell line ab265362 (knockout cell lysate ab257415). To generate this image wild-type and E2F3 knockout HeLa 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 then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-E2F3 antibody (<a href='/en-us/products/primary-antibodies/e2f3-antibody-ab152126'>ab152126</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
E2F3 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human E2F3 knockout HeLa cell line (ab265362)
Lane 3:
SH-SY5Y cell lysate at 20 µg
Lane 4:
U-87 MG cell lysate at 20 µg
Predicted band size: 49 kDa
Observed band size: 49 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human E2F3 knockout HeLa cell line (AB265362)
Allele-2 : 19 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human E2F3 knockout HeLa cell line (AB265362)
Allele-1 : 38 bp deletion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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 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.
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
- 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
DMEM (High Glucose) + 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
E2F3 forms part of the E2F/DP protein complex which controls cell cycle entry and progression. Its activity facilitates the transcription of genes essential for DNA replication and synthesis. Through its interaction with retinoblastoma proteins specifically pRB and p107 E2F3 modulates cellular proliferation. This regulation is vital for maintaining normal cellular functions and growth ensuring proper cell cycle checkpoints are upheld.
Pathways
E2F3 participates in the RB/E2F signaling pathway a major controller of cell proliferation and apoptosis. This pathway serves a critical function in the decision between cell cycle progression and arrest. E2F3 interacts with cyclins and cyclin-dependent kinases (CDKs) particularly CDK2 highlighting its role in the regulation of cell cycle dynamics. It also works alongside other E2F family members such as E2F1 and E2F2 to maintain the balance between cell growth and death.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB320732
Anti-E2F3 antibody [EPR28928-50] - BSA and Azide free
KO Validated|Recombinant|RabMAb
![Western blot - Anti-E2F3 antibody [EPR28928-50] - BSA and Azide free (AB320732)](https://content.abcam.com/products/images/e2f3-antibody-epr28928-50-bsa-and-azide-free-ab320732--western-blot-img435398.jpg)
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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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