Human CDK8 knockout HeLa cell line
- Advanced Validation
- What is this?
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CDK8 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 22 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
CDK8 protein kinase, CDK8_HUMAN, Cell Division Kinase 8, Cell division protein kinase 8, Cyclin-dependent kinase 8, K35, Mediator complex subunit cdk8, Mediator of RNA polymerase II transcription subunit cdk8, Protein kinase K35
- WB
Lab
Western blot - Human CDK8 knockout HeLa cell line (AB265087)
Lanes 1-4 : Merged signal (red and green). Green - ab229192 observed at 53 kDa. Red - loading control ab8245 observed at 37 kDa.
ab229192 Anti-Cdk8 antibody [EPR21005] was shown to specifically react with Cdk8 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265087 (knockout cell lysate ab257885) was used. Wild-type and Cdk8 knockout samples were subjected to SDS-PAGE. ab229192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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-Cdk8 antibody [EPR21005] (<a href='/en-us/products/primary-antibodies/cdk8-antibody-epr21005-ab229192'>ab229192</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDK8 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDK8 knockout HeLa cell line (ab265087)
Lane 3:
HAP1 cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CDK8 knockout HeLa cell line (AB265087)
Allele-1 : 22 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human CDK8 knockout HeLa cell line (AB265087)
Allele-2 : 1 bp insertion in exon 1.
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, 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.
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
Cdk8 plays a critical role in modulating transcriptional responses. It does this by becoming part of the Mediator complex which is essential for transcription regulation by RNA Polymerase II. The Mediator complex where Cdk8 acts as a kinase influences the initiation and regulation of gene expression. Cdk8 phosphorylates several transcription factors and other components affecting their ability to activate or repress transcription.
Pathways
Cyclin-dependent kinase 8 functions prominently in the Wnt/β-catenin signaling and Notch signaling pathways both essential for development and cell differentiation. In the Wnt pathway it interacts closely with proteins like β-catenin to influence gene activation. In the Notch pathway it modulates transcription factors related to cellular differentiation processes. These interactions underline its role as a regulator in carefully managing cellular responses to signaling cues.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Immunoprecipitation - Anti-Cdk8 antibody [EPR21005] (AB229192)](https://content.abcam.com/products/images/cdk8-antibody-epr21005-ab229192--immunoprecipitation-img145440.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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