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Cell Biology Cell Cycle Kinases/Phosphatases Cdks
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Human CDK6 knockout HeLa cell lysate (ab257088)

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Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
  • Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
  • Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
  • Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)
  • Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)

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Overview

  • Product name

    Human CDK6 knockout HeLa cell lysate
    See all Cdk6 kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HeLa
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and Insertion of the selection cassette in exon2.
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    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 - found here. 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.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    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.

    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
    ab260923 - Human CDK6 knockout HeLa cell lysate 1 x 100µg
    ab255929 - Human wild-type HeLa cell lysate 1 x 100µg
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Cdks
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Kinases/Phosphatases
    • Cdks
    • Cancer
    • Cell cycle
    • Kinases/phosphatases
    • Cdks
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Female
  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10

Target

  • Function

    Probably involved in the control of the cell cycle. Interacts with D-type G1 cyclins.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
    Contains 1 protein kinase domain.
  • Target information above from: UniProt accession Q00534 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • CDK 6
    • CDK6
    • CDK6_HUMAN
    • Cell division protein kinase 6
    • Crk 2
    • Crk2
    • Cyclin dependent kinase 6
    • Cyclin-dependent kinase 6
    • MCPH12
    • MGC59692
    • p40
    • PLSTIRE
    • Serine/threonine protein kinase PLSTIRE
    • Serine/threonine-protein kinase PLSTIRE
    • STQTL11
    see all

Associated products

  • KO cell lines

    • Human CDK6 knockout HeLa cell line (ab266059)
  • KO cell pellets

    • Human CDK6 knockout HeLa cell pellet (ab278879)
  • Related Products

    • Anti-Cdk6 antibody [EPR4515] (ab124821)
    • Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
    • Anti-Cdk6 antibody [98D] (ab241554)
    • Anti-Cdk6 antibody [98D] - BSA and Azide free (ab252796)
    • Anti-Cdk6 antibody (ab54576)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab257088 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.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
    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 - ab241554 observed at 38 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
    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 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 ab241554 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
    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 - ab54576 observed at 37 kDa. Red - loading control ab52866 observed at 50 kDa.
    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 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. ab54576 and 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 (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.

  • Western blot - Human CDK6 knockout HeLa cell lysate (ab257088)
    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 - ab124821 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
    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 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. ab124821 and 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 (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 CDK6 knockout HeLa cell lysate (ab257088)
    Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)

    Allele-1: 1 bp insertion in exon2

     

  • Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)
    Sanger Sequencing - Human CDK6 knockout HeLa cell lysate (ab257088)

    Allele-2: Insertion of the selection cassette in exon2

     

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab257088? Please let us know so that we can cite the reference in this datasheet.

ab257088 has not yet been referenced specifically in any publications.

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