CDK4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 139 bp insertion in exon2 and 2 bp deletion 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, 139 bp insertion in exon2 and 2 bp deletion in exon2.
Alternative names
CDK4 protein, CDK4_HUMAN, CMM 3, Cell division kinase 4, Cell division protein kinase 4, Crk3, Cyclin-dependent kinase 4, MGC14458, Melanoma cutaneous malignant 3, PSK-J3, p34 cdk4
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CDK4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 139 bp insertion in exon2 and 2 bp deletion in exon2.
Alternative names
CDK4 protein, CDK4_HUMAN, CMM 3, Cell division kinase 4, Cell division protein kinase 4, Crk3, Cyclin-dependent kinase 4, MGC14458, Melanoma cutaneous malignant 3, PSK-J3, p34 cdk4
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, 139 bp insertion in exon2 and 2 bp deletion in exon2.
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
CDK4
- 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
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Cdk4 also known as Cyclin-dependent kinase 4 is a serine/threonine kinase involved in cell cycle regulation. Cdk4 has a molecular weight of approximately 34 kDa. This protein mainly locates in the nucleus of the cell and its expression occurs in diverse tissues. Cdk4 partners with D-type cyclins to exert its function which is essential for the transition from the G1 phase to the S phase of the cell cycle. Researchers often study Cdk4 using techniques like immunohistochemistry (IHC) to determine its expression and localization in different tissues.
- Biological function summary
Cdk4 plays a fundamental role in cell cycle regulation by forming a complex with D-type cyclins. Together they phosphorylate the retinoblastoma protein (Rb) resulting in the release of E2F transcription factors which promote the expression of genes necessary for DNA replication. The Cdk4/cyclin D complex regulates the cell's commitment to enter S phase and continue cell division. This regulation is key for normal cell proliferation and tissue homeostasis. Cdk4 activity is strictly controlled by INK4 family members acting as inhibitors and adding an extra regulation layer.
- Pathways
Cdk4 is an integral part of important signaling pathways like the cell cycle and PI3K/AKT pathways. In the cell cycle pathway Cdk4 relays signals downstream that drive the transition from the G1 to S phase through its interaction with cyclin D and Rb. In the PI3K/AKT pathway signaling can influence cyclin D expression indirectly affecting Cdk4 activity. Proteins such as Cdk6 closely relate to Cdk4 and often compensate or partner with Cdk4 in these pathways to maintain proper cell cycle progression.
- Associated diseases and disorders
Abnormalities in Cdk4 function or regulation relate to cancer and certain metabolic syndromes. Hyperactivity of Cdk4 due to overexpression or mutation frequently occurs in several cancers including melanoma and breast cancer. In these malignancies Cdk4 aberrantly drives excessive cell proliferation. Mutations or alterations in proteins such as p16 an inhibitor of Cdk4 are often found in these cancers highlighting the critical role of Cdk4 in disease progression. Understanding the role of Cdk4 allows researchers to develop targeted therapies such as Cdk4/6 inhibitors offering new treatment options.
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3 product images
Western blot - Human CDK4 knockout HeLa cell lysate (ab263780)
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: CDK4 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cdk4 antibody [EPR4513-32-7] ab108357 observed at 34 kDa. Red - loading control Anti-Vinculin antibody [VIN-54] ab130007 observed at 124 kDa.
Anti-Cdk4 antibody [EPR4513-32-7] ab108357 Recombinant Anti-Cdk4 antibody [EPR4513-32-7] was shown to specifically react with CDK4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK4 knockout HeLa cell line ab255378 (knockout cell lysate ab263780) was used. Wild-type and CDK4 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-Cdk4 antibody [EPR4513-32-7] ab108357 and Anti-Vinculin antibody [VIN-54] 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-Cdk4 antibody [EPR4513-32-7] (Anti-Cdk4 antibody [EPR4513-32-7] ab108357) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CDK4 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa, 55 kDa
Observed band size: 34 kDa, 55 kDa
Western blot - Human CDK4 knockout HeLa cell lysate (ab263780)
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: CDK4 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cdk4 antibody [EPR17525] ab199728 observed at 34 kDa. Red - loading control Anti-Vinculin antibody [VIN-54] ab130007 observed at 124 kDa.
Anti-Cdk4 antibody [EPR17525] ab199728 Recombinant Anti-Cdk4 antibody [EPR17525] was shown to specifically react with CDK4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK4 knockout HeLa cell line ab255378 (knockout cell lysate ab263780) was used. Wild-type and CDK4 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-Cdk4 antibody [EPR17525] ab199728 and Anti-Vinculin antibody [VIN-54] were incubated overnight at 4°C at 1 in 2000 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-Cdk4 antibody [EPR17525] (Anti-Cdk4 antibody [EPR17525] ab199728) at 1/2000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CDK4 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Sanger Sequencing - Human CDK4 knockout HeLa cell lysate (ab263780)
Allele-1: 2 bp deletion in exon2; Allele-2: 139 bp insertion in exon2
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