Astana Biomed Group, an authorized Abcam distributor in Central Asia

Human GLYR1 knockout HEK-293T cell lysate (ab257968)

Overview

  • Product name

    Human GLYR1 knockout HEK-293T cell lysate

  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp insertion in exon 3.

  • Passage number

  • 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

Target

  • Function

    May have oxidoreductase activity. Regulates p38 MAP kinase activity by mediating stress activation of p38alpha/MAPK14 and specifically regulating MAPK14 signaling. Indirectly promotes phosphorylation of MAPK14 and activation of ATF2. The phosphorylation of MAPK14 requires upstream activity of MAP2K4 and MAP2K6. Recruited on chromatin, recognizes and binds trimethylated 'Lys-36' of histone H3 (H3K36me3).

  • Sequence similarities

    Belongs to the 3-hydroxyisobutyrate dehydrogenase family. NP60 subfamily.
    Contains 1 A.T hook DNA-binding domain.
    Contains 1 PWWP domain.

  • Domain

    The A.T hook DNA-binding domain is required for the interaction with MAPK14.
    The PWWP domain probably mediates the binding to H3K36me3.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Alternative names

    • 3 hydroxyisobutyrate dehydrogenase like protein
    • 3-hydroxyisobutyrate dehydrogenase-like protein
    • BM045
    • Cytokine like nuclear factor n pac
    • Cytokine-like nuclear factor N-PAC
    • Glyoxylate reductase 1 homolog
    • GLYR1
    • GLYR1_HUMAN
    • HIBDL
    • N-PAC
    • NP60
    • Nuclear protein 60 kDa
    • Nuclear protein NP60
    • Nuclear protein of 60 kDa
    • Putative oxidoreductase GLYR1
    see all

Properties

Images

  • Western blot - Human GLYR1 knockout HEK293T cell lysate (ab257968)
    Western blot - Human GLYR1 knockout HEK293T cell lysate (ab257968)

    Lane 1:Wild-type HEK293T cell lysate (20 ug)
    Lane 2:GLYR1 knockout HEK293T cell lysate (20 ug)
    Lane 3:HeLa cell lysate (20 ug)
    Lane 4:HUVEC cell lysate (20 ug)

    ab167155 was shown to specifically react with GLYR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266610 (knockout cell lysate ab257968) was used. Wild-type and GLYR1 knockout samples were subjected to SDS-PAGE. ab167155 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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.

  • Western blot - Human GLYR1 knockout HEK293T cell lysate (ab257968)
    Western blot - Human GLYR1 knockout HEK293T cell lysate (ab257968)

    Lane 1:Wild-type HEK293T cell lysate (20 ug)
    Lane 2:GLYR1 knockout HEK293T cell lysate (20 ug)
    Lane 3:HeLa cell lysate (20 ug)
    Lane 4:HUVEC cell lysate (20 ug)

    ab154838 was shown to specifically react with GLYR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266610 (knockout cell lysate ab257968) was used. Wild-type and GLYR1 knockout samples were subjected to SDS-PAGE. ab154838 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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 GLYR1 knockout HEK293T cell lysate (ab257968)
    Sanger Sequencing - Human GLYR1 knockout HEK293T cell lysate (ab257968)
    Homozygous: 10 bp insertion in exon 3

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Clear all

Recently viewed products

Sign up