NEK2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 53 bp deletion in exon 1.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 53 bp deletion in exon 1
Recommended products
NEK2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 53 bp deletion in exon 1.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 53 bp deletion in exon 1
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
NEK2
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Cell culture
- Biosafety level
EU: 2 US: 2
- Viability
~ 80%
- Adherent/suspension
Adherent
- Gender
Female
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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab271142). 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: DMEM (High Glucose) + 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 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 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
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 cell cycle serine/threonine-protein kinase Nek2 also known as Never in Mitosis Gene A (NIMA)-related kinase 2 has a molecular weight of approximately 56 kDa. Expressed mainly in proliferating cells with high expression in the testes it primarily localizes to the centrosome and nucleus. This protein regulates centrosome separation and spindle formation. Such functions are essential for proper cell division contributing to the accurate segregation of chromosomes.
- Biological function summary
Nek2 involves key roles in cell cycle progression especially from the G2 to the M phase. It integrates into a complex regulatory network interacting with various cell division controllers. Nek2 phosphorylates centrosomal proteins which ensures timely centrosome disjunction and bipolar spindle formation. This action establishes Nek2 as important for the maintenance of genomic stability preventing abnormal cell divisions.
- Pathways
Nek2 fits into critical pathways that control cell division and mitosis. It operates within the PI3K/AKT pathway influencing cell growth and survival. Additionally Nek2 relates to other kinases like Aurora A kinase sharing pathway intersections that modulate centrosome dynamics and stability. These interactions reflect the significance of Nek2 in orchestrating precise mitotic events contributing to orderly cell cycle transitions.
- Associated diseases and disorders
Nek2 shows connections to cancers particularly breast and prostate cancer. Overexpression of Nek2 correlates with the pathogenesis of these cancers indicating its role in aberrant cellular proliferation. It also interacts with proteins like MAD2 which is a spindle checkpoint protein. This connection highlights Nek2's involvement in tumorigenesis through mitotic checkpoint control failure contributing to cancer cell survival and progression.
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1 product image
Sanger Sequencing - Human NEK2 knockout HeLa cell line (ab266027)
Homozygous: 53 bp deletion in exon1
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