Overview

  • Product name

    Human KRT8 knockout HeLa cell lysate
  • Product overview


    Cell line information
    Parental cell line: HeLa
    Organism: Homo sapiens

    Gene editing information
    Editing tool: CRISPR/Cas9
    Mutation: 1 bp insertion in exon 2 and 2 bp deletion in exon 2 and 4 bp deletion in exon 2.

    Knockout validation: Confirmed by Sanger sequencing and WB.

    Reconstitution instructions: To use as a WB control, resuspend in 45 µL of Sample buffer (40% (w/v) Glycerol, 4% (w/v) Lithium Dodecyl Sulfate, 4% Ficoll 400, 0.025% Phenol Red, 0.025% Brilliant Blue G250, 2 mM EDTA) and 5 µL of DTT to resuspend @ 2mg/ml. Mix well, then boil the sample for 10 minutes before loading it onto the gel.

    User storage instructions: Upon receiving, lysate can be diluted with 1 x SDS sample buffer & will be stable at -20°C for 12 months. Long term storage at -80°C.

  • Notes

    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

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    Human KRT8 knockout HeLa cell lysate (Lyophilized) 1 x 100µg
    Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
  • Research areas

  • Function

    Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.
  • Tissue specificity

    Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma membrane in structures that contain dystrophin and spectrin. Expressed in gingival mucosa and hard palate of the oral cavity.
  • Involvement in disease

    Cirrhosis
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Post-translational
    modifications

    Phosphorylation on serine residues is enhanced during EGF stimulation and mitosis. Ser-74 phosphorylation plays an important role in keratin filament reorganization.
    O-glycosylated. O-GlcNAcylation at multiple sites increases solubility, and decreases stability by inducing proteasomal degradation.
    O-glycosylated (O-GlcNAcylated), in a cell cycle-dependent manner.
  • Cellular localization

    Cytoplasm. Nucleus, nucleoplasm. Nucleus matrix.
  • Information by UniProt
  • Alternative names

    • CARD2
    • CK 8
    • CK-8
    • CK8
    • CYK8
    • CYKER
    • Cytokeratin endo A
    • Cytokeratin-8
    • DreK8
    • EndoA
    • K2C8
    • K2C8_HUMAN
    • K8
    • Keratin 8
    • Keratin type II cytoskeletal 8
    • Keratin, type II cytoskeletal 8
    • Keratin-8
    • KO
    • Krt 2.8
    • KRT8
    • MGC118110
    • MGC174782
    • MGC53564
    • MGC85764
    • sb:cb186
    • Type-II keratin Kb8
    see all

Applications

Our Abpromise guarantee covers the use of ab263785 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration.

Images

  • Lane 1: A431 cell lysate (20 µg)
    Lane 2: MCF7 cell lysate (20 µg)
    Lane 3: HeLa wildtype cell lysate (20 µg)
    Lane 4: KRT8 HeLa knockout cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab9023 observed at 55 kDa. Red - loading control, ab181602 observed at 37 kDa.


    ab9023 was shown to react with Cytokeratin 8 in HeLa wildtype. Loss of signal was observed when knockout sample ab263785 was used. Wild-type and Cytokeratin 8 knockout samples were subjected to SDS-PAGE. ab9023 and Anti-GAPDH antibody EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and 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.

  • Lane 1: A431 cell lysate (20 µg)
    Lane 2: MCF7 cell lysate (20 µg)
    Lane 3: HeLa wildtype cell lysate (20 µg)
    Lane 4: KRT8 HeLa knockout cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab53280 observed at 55 kDa. Red - loading control, ab8245 observed at 37 kDa.


    ab53280 was shown to react with Cytokeratin 8 in HeLa wildtype. Loss of signal was observed when knockout sample ab263785 was used. Wild-type and Cytokeratin 8 knockout samples were subjected to SDS-PAGE. ab53280 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 (For unpurified use at 1/25,000 - 1/50,000) 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.

  • Allel-1: 4 bp deletion in exon 2
  • Allel-2: 2 bp deletion in exon 2
  • Allel-3: 1 bp insertion in exon 2

References

ab263785 has not yet been referenced specifically in any publications.

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