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AB262513

Human CENPF knockout HeLa cell line

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CENPF KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 20 bp deletion after Phe38 of the WT protein Frameshift: 98.64%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Next Generation Sequencing - Human CENPF knockout HeLa cell line (AB262513)
  • NGS

Lab

Next Generation Sequencing - Human CENPF knockout HeLa cell line (AB262513)

20 bp deletion after Phe38 of the WT protein

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Mutation description

Knockout achieved by CRISPR/Cas9 X = 20 bp deletion after Phe38 of the WT protein Frameshift: 98.64%

Disease

Adenocarcinoma

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.

What's included?

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Properties and storage information

Gene name
CENPF
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

CENPF also known as centromere protein F or mitosin is a significant kinetochore-associated protein. It has a molecular mass of approximately 367 kDa. Expressed chiefly in the nucleus CENPF displays elevated levels during cell division particularly mitosis. The protein accumulates at the nuclear matrix during the G2 phase of the cell cycle and moves to the kinetochores in mitosis highlighting its role in cell cycle regulation.
Biological function summary

It plays a role in chromosome segregation and spindle attachment during cell division. CENPF associates with various cellular complexes including the nuclear matrix and kinetochores. Its function is critical to maintaining stability and proper chromosome movement during mitosis. This involvement ensures accurate chromosomal alignment and segregation preventing genomic instability in daughter cells.

Pathways

CENPF plays an essential role in the regulation of the mitotic cell cycle and chromosomal dynamics. The protein interfaces with several elements in pathways like the spindle assembly checkpoint. It interacts notably with proteins such as CENP-E and other kinetochore constituents. These interactions safeguard the precise attachment of microtubules to the kinetochores facilitating proper cell cycle progression and division.

Alterations in CENPF expression and function have been linked to cancer and congenital heart defects. Dysregulation of CENPF can lead to chromosomal instability a known hallmark of many cancers. Additionally its interactions with proteins like CENP-E and Bub1 further relate to its involvement in the proper execution of cell cycle checkpoints. Such associations make CENPF a potential target for therapeutic interventions in related disorders.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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