CDK6 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
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 lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
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
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
- 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
Storage
- Shipped at conditions
Ambient - Can Ship with Ice
- Appropriate long-term storage conditions
-20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
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5 product images
Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
Lane 1: Wild-type HeLa cell lysate 20 ug
Lane 2: CDK6 knockout HeLa cell lysate 20 ug
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 55kDa.
Anti-Cdk6 antibody [98D] ab241554 was shown to react with Cdk6 in wild-type HeLa cells in Western blot with loss of signal observed in CDK6 knockout cell line Human CDK6 knockout HeLa cell line ab266059 (CDK6 knockout 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 °C 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
Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
Lane 1: Wild-type HeLa cell lysate (20μg)
Lane 2: CDK6 knockout HeLa cell lysate (20μg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cdk6 antibody [EPR4515] ab124821 observed at 37 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.
Anti-Cdk6 antibody [EPR4515] ab124821 Anti-Cdk6 antibody [EPR4515] was shown to specifically react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK6 knockout HeLa cell line ab266059 (knockout cell lysate ab257088) was used. Wild-type and Cdk6 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Cdk6 antibody [EPR4515] ab124821 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-Cdk6 antibody [EPR4515] (Anti-Cdk6 antibody [EPR4515] ab124821) at 1/1000 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: 37 kDa
Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
Lane 1: Wild-type HeLa cell lysate (20μg)
Lane 2: CDK6 knockout HeLa cell lysate (20μg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cdk6 antibody [8H4] ab54576 observed at 37 kDa. Red - loading control Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866 observed at 50 kDa.
Anti-Cdk6 antibody [8H4] ab54576 Anti-Cdk6 antibody was shown to specifically react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK6 knockout HeLa cell line ab266059 (knockout cell lysate ab257088) was used. Wild-type and Cdk6 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Cdk6 antibody [8H4] ab54576 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) were incubated overnight at 4°C at 1 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-Cdk6 antibody [8H4] (Anti-Cdk6 antibody [8H4] ab54576)
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: 37 kDa
Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)
Allele-2: Insertion of the selection cassette in exon2
Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)
Allele-1: 1 bp insertion in exon2
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