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E2F1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.

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Images

Western blot - Human E2F1 knockout HCT116 cell line (AB287380), expandable thumbnail
  • Western blot - Human E2F1 knockout HCT116 cell line (AB287380), expandable thumbnail
  • Next Generation Sequencing - Human E2F1 knockout HCT116 cell line (AB287380), expandable thumbnail

Key facts

Cell type
HCT116
Species or organism
Human
Tissue
Colon
Form
Liquid
Knockout validation
Western blot

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

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E2F1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.

Key facts

Cell type
HCT116
Form
Liquid
Disease
Carcinoma
Concentration
Loading...

Properties

Gene name
E2F1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Western blot

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
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 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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HCT116 cell line (ab288559). 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.


Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.


Culture medium:  McCoY5a + 10% FBS


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. Based on cell count, seed cells in an appropriate cell culture 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.
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 2x104 cells/cm2 is recommended.
  • A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
  • Cells should be passaged when they have achieved 80-90% confluence.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The E2F1 protein also known as E2F transcription factor 1 is an important regulator of the cell cycle. It has a molecular weight of approximately 47 kDa. E2F1 protein is expressed in many tissues with higher expression levels in proliferating cells. It often acts as a transcription factor by binding to specific DNA sequences controlling the transition of cells from G1 to S phase of the cell cycle. This role is vital for the proper progression of the cycle and cellular proliferation.

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.

Associated diseases and disorders

E2F1 is often linked to cancer and neurodegenerative diseases. Overexpression or dysregulation of E2F1 can lead to uncontrolled cell proliferation contributing to oncogenesis. In the context of neurodegenerative diseases E2F1 influences neuronal apoptosis and may be associated with disorders such as Alzheimer's. E2F1 interacts with other proteins such as p53 in cancer where it affects both cell cycle dynamics and apoptosis illustrating its complex involvement in disease progression.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Western blot - Human E2F1 knockout HCT116 cell line (ab287380), expandable thumbnail

    Western blot - Human E2F1 knockout HCT116 cell line (ab287380)

    Western blot: Anti-E2F1 antibody [EPR26698-49] (Anti-E2F1 antibody [EPR26698-49] ab314311) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-E2F1 antibody [EPR26698-49] 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] (Anti-E2F1 antibody [EPR26698-49] ab314311) 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 Human E2F1 knockout HEK-293 cell line ab269502 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

    Performed under reducing conditions.

  • Western blot - Human E2F1 knockout HCT116 cell line (ab287380), expandable thumbnail

    Western blot - Human E2F1 knockout HCT116 cell line (ab287380)

    Western blot: Anti-E2F1 antibody [KH95] (Anti-E2F1 antibody [KH95] ab4070) staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-E2F1 antibody [KH95] 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] (Anti-E2F1 antibody [KH95] ab4070) 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 Human E2F1 knockout HEK-293 cell line ab269502 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

    Performed under reducing conditions.

  • Next Generation Sequencing - Human E2F1 knockout HCT116 cell line (ab287380), expandable thumbnail

    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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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