Overview

  • Product name
    Anti-ATRX antibody [CL0537]
    See all ATRX primary antibodies
  • Description
    Mouse monoclonal [CL0537] to ATRX
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Recombinant fragment corresponding to Human ATRX aa 2273-2413.
    Sequence:

    AAWAEYEAEKKGLTMRFNIPTGTNLPPVSFNSQTPYIPFNLGALSAMSNQ QLEDLINQGREKVVEATNSVTAVRIQPLEDIISAVWKENMNLSEAQVQAL ALSRQASQELDVKRREAIYNDVLTKQQMLISCVQRILMNRR


    Database link: P46100

  • Positive control
    • Human brain glioma, fallopian tube, kidney, prostate, stomach, colorectal cancer, cerebellum and hippocampus tissue WB: A-549 cells

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.2
    Preservative: 0.02% Sodium azide
    Constituents: 59% PBS, 40% Glycerol
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    CL0537
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab188027 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
IHC-P 1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/500 - 1/1000. Predicted molecular weight: 282 kDa.
ICC/IF Use a concentration of 1 - 4 µg/ml.

Fixation/Permeabilization: PFA/Triton X-100.

Target

  • Function
    Involved in transcriptional regulation and chromatin remodeling. Facilitates DNA replication in multiple cellular environments and is required for efficient replication of a subset of genomic loci. Binds to DNA tandem repeat sequences in both telomeres and euchromatin and in vitro binds DNA quadruplex structures. May help stabilizing G-rich regions into regular chromatin structures by remodeling G4 DNA and incorporating H3.3-containing nucleosomes. Catalytic component of the chromatin remodeling complex ATRX:DAXX which has ATP-dependent DNA translocase activity and catalyzes the replication-independent deposition of histone H3.3 in pericentric DNA repeats outside S-phase and telomeres, and the in vitro remodeling of H3.3-containing nucleosomes. Its heterochromatin targeting is proposed to involve a combinatorial readout of histone H3 modifications (specifically methylation states of H3K9 and H3K4) and association with CBX5. Involved in maintaining telomere structural integrity in embryonic stem cells which probably implies recruitment of CBX5 to telomers. Reports on the involvement in transcriptional regulation of telomeric repeat-containing RNA (TERRA) are conflicting; according to a report, it is not sufficient to decrease chromatin condensation at telomers nor to increase expression of telomeric RNA in fibroblasts (PubMed:24500201). May be involved in telomere maintenance via recombination in ALT (alternative lengthening of telomeres) cell lines. Acts as negative regulator of chromatin incorporation of transcriptionally repressive histone H2AFY, particularily at telomeres and the alpha-globin cluster in erythroleukemic cells. Participates in the allele-specific gene expression at the imprinted IGF2/H19 gene locus. On the maternal allele, required for the chromatin occupancy of SMC1 and CTCTF within the H19 imprinting control region (ICR) and involved in esatblishment of histone tails modifications in the ICR. May be involved in brain development and facial morphogenesis. Binds to zinc-finger coding genes with atypical chromatin signatures and regulates its H3K9me3 levels. Forms a complex with ZNF274, TRIM28 and SETDB1 to facilitate the deposition and maintenance of H3K9me3 at the 3' exons of zinc-finger genes (PubMed:27029610).
  • Tissue specificity
    Ubiquitous.
  • Involvement in disease
    Alpha-thalassemia mental retardation syndrome, X-linked
    Mental retardation, X-linked, syndromic, with hypotonic facies 1
    Alpha-thalassemia myelodysplasia syndrome
  • Sequence similarities
    Belongs to the SNF2/RAD54 helicase family.
    Contains 1 ADD domain.
    Contains 1 GATA-type zinc finger.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 1 PHD-type zinc finger.
  • Domain
    The ADD domain predominantly interacts with histone H3 trimethylated at 'Lys-10'(H3K9me3) (and to a lesser extent H3 mono-or dimethylated at 'Lys-10') and simultanously to histone H3 unmethylated at 'Lys-5' (H3K4me0). The interaction with H3K9me3 is disrupted by the presence of H3K4me3 suggesting a readout of the combined histone H3 methylation state.
    Contains one Pro-Xaa-Val-Xaa-Leu (PxVxL) motif, which is required for interaction with chromoshadow domains. This motif requires additional residues -7, -6, +4 and +5 of the central Val which contact the chromoshadow domain.
  • Post-translational
    modifications
    Phosphorylated at serine residues during mitose. Phosphorylation may promote the release from the nuclear matrix and progression to mitosis.
  • Cellular localization
    Nucleus. Chromosome, telomere. Nucleus, PML body. Associated with pericentromeric heterochromatin during interphase and mitosis, probably by interacting with CBX5/HP1 alpha. Colocalizes with histone H3.3, DAXX, HIRA and ASF1A at PML-nuclear bodies. Colocalizes with cohesin (SMC1 and SMC3) and MECP2 at the maternal H19 ICR (By similarity).
  • Information by UniProt
  • Database links
  • Alternative names
    • Alpha thalassemia/mental retardation syndrome X linked homolog antibody
    • ATP dependent helicase ATRX antibody
    • ATP-dependent helicase ATRX antibody
    • ATR2 antibody
    • Atrx antibody
    • ATRX_HUMAN antibody
    • DNA dependent ATPase and helicase antibody
    • Helicase 2, X linked antibody
    • MGC2094 antibody
    • MRXHF1 antibody
    • RAD54 antibody
    • RAD54L antibody
    • SFM1 antibody
    • SHS antibody
    • Transcriptional regulator ATRX antibody
    • X linked helicase II antibody
    • X linked nuclear protein antibody
    • X-linked helicase II antibody
    • X-linked nuclear protein antibody
    • XH2 antibody
    • XNP antibody
    • Znf HX antibody
    • Znf-HX antibody
    see all

Images

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human cerebellum tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling ATRX with ab188027 showing clear nuclear (without nucleoli) staining in green. Microtubule are visualized in red.



  • Predicted band size : 282 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: ATRX knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab188027 observed at 300 kDa. Red - loading control, ab176560, observed at 50 kDa.

    ab188027 was shown to recognize ATRX in wild type cells as signal was lost at the expected MW in ATRX knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ATRX knockout samples were subjected to SDS-PAGE. Ab188027 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at a 1/500 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

     

  • All lanes : Anti-ATRX antibody [CL0537] (ab188027)

    Lane 1 : A-549 cells transfected with control siRNA
    Lane 2 : A-549 cells transfected with target specific siRNA probe #1
    Lane 3 : A-549 cells transfected with target specific siRNA probe #2


    Predicted band size : 282 kDa

    Downregulation of antibody signal confirms target specificity. Remaining % intensity, relative control lane, is indicated. Anti-GAPDH monoclonal antibody was used as loading control.

  • Lane 1 :
    Lane 2 : Anti-ATRX antibody [CL0537] (ab188027) at 1/500 dilution

    Lane 1 : Marker [kDa]
    Lane 2 : Human cell line A-549


    Predicted band size : 282 kDa
  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human fallopian tube tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human brain (high grade glioma) tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human hippocampus tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human prostate tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human stomach tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colorectal cancer tissue, labeling ATRX with ab188027 at 1/200 dilution.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ATRX with ab188027 showing clear nuclear (without nucleoli) staining in green. Microtubule are visualized in red.

References

ab188027 has not yet been referenced specifically in any publications.

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