Human NEK9 knockout A-431 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human NEK9 knockout A-431 cell line (AB270474)
Lanes 1 - 4 : Merged signal (red and green). Green - ab138488 observed at 120 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
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 ab138488 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-NEK9 antibody [EP7361] (<a href='/en-us/products/primary-antibodies/nek9-antibody-ep7361-ab138488'>ab138488</a>) 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
Predicted band size: 107 kDa
Observed band size: 120 kDa
false
- NGS
Supplier Data
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
- NGS
Supplier Data
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%
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Cell Lines & Lysates
AB267247
Human EIF2AK4 (GCN2) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
Reactivity data
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?
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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