NEK9 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 10 bp deletion, 2 bp deletion; Frameshift: 99.78%.
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Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 10 bp deletion, 2 bp deletion; Frameshift: 99.78%
Alternative names
APUG, DKFZp434D0935, KIAA1995, LCCS10, MGC138306, MGC16714, NC, NEK9_HUMAN, NERCC, NERCC1, NIMA (never in mitosis gene a) related kinase 9, NIMA related kinase 9, NIMA related kinase Nek8, NIMA-related kinase 8, Nercc 1 kinase, Never in mitosis A-related kinase 9, NimA-related protein kinase 9, Serine/threonine-protein kinase Nek9
Recommended products
NEK9 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 10 bp deletion, 2 bp deletion; Frameshift: 99.78%.
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 10 bp deletion, 2 bp deletion; Frameshift: 99.78%
- Disease
- Epidermoid Carcinoma
- Concentration
Properties
- Gene name
- NEK9
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, 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
- 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
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
NEK9 also known as NIMA-related kinase 9 is a serine/threonine kinase with a molecular mass of approximately 118 kDa. It plays a role in the regulation of various cell cycle processes. NEK9 is active in both the cytoplasm and nucleus and expresses broadly in various tissues including the liver kidneys and pancreas. It also has homologs in other eukaryotic organisms indicating its conserved function across species.
- Biological function summary
This protein modulates different aspects of mitosis particularly in spindle formation and chromosome segregation. NEK9 forms a part of a complex in conjunction with other kinases like NEK6 and NEK7 coordinating the early events of mitosis. Its activity supports centrosome separation and microtubule nucleation which are important for proper cell division.
- Pathways
NEK9 is involved in the cell cycle and microtubule organization pathways. It works in conjunction with proteins such as PLK1 and Aurora A which regulate the entry and progression through mitosis. NEK9 activates NEK6 and NEK7 which further propagate signals necessary for mitotic spindle assembly ensuring efficient cell division.
- Associated diseases and disorders
NEK9 has links to cancer and neurological disorders. Abnormal NEK9 activity has associations with tumorigenesis particularly linked with the dysregulation of cell cycle checkpoints and malignant transformation. In neurological disorder contexts NEK9 interacts with proteins like tau perturbing normal cellular functions and potentially contributing to disease pathology.
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3 product images
Western blot - Human NEK9 knockout A-431 cell line (ab270474)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-NEK9 antibody [EP7361] ab138488 observed at 120 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.Anti-NEK9 antibody [EP7361] ab138488 was shown to react with NEK9 in wild-type A431 cells in western blot. Loss of signal was observed when NEK9 knockout sample was used. Wild-type and NEK9 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-NEK9 antibody [EP7361] ab138488 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-NEK9 antibody [EP7361] (Anti-NEK9 antibody [EP7361] ab138488) at 1/1000 dilution
Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2: NEK9 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human NEK9 knockout A-431 cell line (ab270474)
Lane 3: Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 107 kDa
Observed band size: 120 kDa
Next Generation Sequencing - Human NEK9 knockout A-431 cell line (ab270474)
Knockout achieved by CRISPR/Cas9; X = 10 bp deletion, 2 bp deletion; Frameshift: 99.78%
Next Generation Sequencing - Human NEK9 knockout A-431 cell line (ab270474)
10 bp deletion after Glu81 (allele 1) and 2 bp deletion after Asp83 (allele 2) of the WT protein
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