CDK6 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2
Alternative names
CDK6_HUMAN, Cell division protein kinase 6, Crk 2, Cyclin-dependent kinase 6, MCPH12, MGC59692, PLSTIRE, STQTL11, Serine/threonine-protein kinase PLSTIRE
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CDK6 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Alternative names
CDK6_HUMAN, Cell division protein kinase 6, Crk 2, Cyclin-dependent kinase 6, MCPH12, MGC59692, PLSTIRE, STQTL11, Serine/threonine-protein kinase PLSTIRE
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, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
CDK6
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- 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 long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255928). 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.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Supplementary info
- Activity summary
Cdk6 also known as Cyclin-dependent kinase 6 is a protein kinase involved in the regulation of the cell cycle. It has a molecular weight of approximately 40 kDa. Cdk6 is expressed in various tissues including lymphoid tissues and the brain. It plays a mechanical role by phosphorylating target proteins which impacts cell cycle progression particularly during the transition from G1 phase to S phase. Cdk6 can work in conjunction with D-type cyclins to form a complex that is critical for its activity.
- Biological function summary
Cdk6 functions as a regulator of the cell cycle. It participates in a complex with cyclin D to control the G1 phase progression. This activity is essential for the regulation of cell division and proliferation. The association with cyclin D allows Cdk6 to phosphorylate the retinoblastoma protein (Rb) leading to the release of E2F transcription factors that promote the expression of genes necessary for DNA synthesis.
- Pathways
Cdk6 is an important component of the cell cycle control pathway. It interacts primarily with the retinoblastoma protein and Cyclin D1 in this context. The cell cycle pathway is critical for controlled cell proliferation and its dysregulation can lead to diseases like cancer. Another involved pathway is the PI3K/AKT pathway which can activate Cdk6 activity through upstream signaling events implicating Cdk6 in cellular responses to growth signals.
- Associated diseases and disorders
Cdk6 has a significant role in cancer development due to its involvement in uncontrolled cell proliferation. Its overexpression or dysregulation is often linked to cancers such as leukemia and glioblastoma. In cancer the interaction with proteins like cyclin D1 can lead to unchecked progression through the cell cycle contributing to oncogenesis. Additionally Cdk6 inhibitors are being explored as therapeutic agents in cancer treatment due to their potential to restore control over cell cycle progression.
Product promise
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4 product images
Western blot - Human CDK6 knockout HeLa cell line (ab266059)
Lanes 1 - 2: Merged signal (red and green). Green - Anti-Cdk6 antibody [98D] ab241554 observed at 38 kDa. Red - loading control Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.
Anti-Cdk6 antibody [98D] ab241554 was shown to react with Cdk6 in HeLa wild-type cells in Western blot with loss of signal observed in CDK6 knockout cell line ab266059 (CDK6 knockout cell lysate Human CDK6 knockout HeLa cell lysate ab257088). Wild-type HeLa and CDK6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-Cdk6 antibody [98D] ab241554 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4° at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-Cdk6 antibody [98D] (Anti-Cdk6 antibody [98D] ab241554) at 1/2000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CDK6 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 38 kDa
Cell Culture - Human CDK6 knockout HeLa cell line (ab266059)
Representative images of CDK6 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human CDK6 knockout HeLa cell line (ab266059)
Allele-2: Insertion of the selection cassette in exon 2.
Sanger Sequencing - Human CDK6 knockout HeLa cell line (ab266059)
Allele-1: 1 bp insertion in exon2
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