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Human CDK8 knockout HeLa cell pellet (ab278917)

Overview

  • Product name

    Human CDK8 knockout HeLa cell pellet
    See all Cdk8 kits

  • Product overview

    Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.

    Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation.

  • Parental Cell Line

    HeLa

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and 22 bp deletion in exon1.

  • Passage number

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)

  • Notes

    Pellet size: 5 million cells/vial.

    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.

  • Tested applications

    Suitable for: WBmore details

Properties

Target

  • Function

    Component of the Mediator complex, a coactivator involved in regulated gene transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. Phosphorylates the CTD (C-terminal domain) of the large subunit of RNA polymerase II (RNAp II), which may inhibit the formation of a transcription initiation complex. Phosphorylates CCNH leading to down-regulation of the TFIIH complex and transcriptional repression. Recruited through interaction with MAML1 to hyperphosphorylate the intracellular domain of NOTCH, leading to its degradation.

  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
    Contains 1 protein kinase domain.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Alternative names

    • cdk8
    • CDK8 protein kinase
    • CDK8_HUMAN
    • Cell Division Kinase 8
    • Cell division protein kinase 8
    • Cyclin Dependent kinase 8
    • Cyclin-dependent kinase 8
    • K35
    • Mediator complex subunit cdk8
    • Mediator of RNA polymerase II transcription subunit cdk8
    • Protein kinase K35
    see all

Properties

Images

  • Western blot - Human CDK8 knockout HeLa cell pellet (ab278917)
    Western blot - Human CDK8 knockout HeLa cell pellet (ab278917)

    Lane 1: Wild-type HeLa cell lysate (20 µg)

    Lane 2: CDK8 knockout HeLa cell lysate (20 µg)

    Lane 3: HAP1 cell lysate (20 µg)

    Lane 4: Daudi cell lysate (20 µg)

    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°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 (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.

  • Sanger Sequencing - Human CDK8 knockout HeLa cell pellet (ab278917)
    Sanger Sequencing - Human CDK8 knockout HeLa cell pellet (ab278917)

    Allele-1: 22 bp deletion in exon1

     

  • Sanger Sequencing - Human CDK8 knockout HeLa cell pellet (ab278917)
    Sanger Sequencing - Human CDK8 knockout HeLa cell pellet (ab278917)

    Allele-2: 1 bp insertion in exon1

     

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