Anti-ATRX antibody [CL0537] (ab188027)
Key features and details
- Mouse monoclonal [CL0537] to ATRX
- Suitable for: IHC-P, ICC, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-ATRX antibody [CL0537]
See all ATRX primary antibodies -
Description
Mouse monoclonal [CL0537] to ATRX -
Host species
Mouse -
Tested applications
Suitable for: IHC-P, ICC, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Recombinant fragment corresponding to Human ATRX aa 2250-2450.
Database link: P46100 -
Positive control
- IHC-P: Human brain glioma, fallopian tube, and testis tissues; WB: A-549 cell lysate. ICC: A431 and HeLa cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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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: 7.20
Preservative: 0.02% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
CL0537 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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IHC-P |
1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC |
Use a concentration of 2 - 10 µg/ml.
Fixation/Permeabilization: PFA/Triton X-100. |
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WB |
Use a concentration of 1 µg/ml. Predicted molecular weight: 282 kDa.
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Notes |
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IHC-P
1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC
Use a concentration of 2 - 10 µg/ml. Fixation/Permeabilization: PFA/Triton X-100. |
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 282 kDa. |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 546 Human
- Entrez Gene: 22589 Mouse
- Entrez Gene: 246284 Rat
- Omim: 301040 Human
- SwissProt: P46100 Human
- SwissProt: Q61687 Mouse
- SwissProt: P70486 Rat
- Unigene: 533526 Human
see all -
Alternative names
- Alpha thalassemia/mental retardation syndrome X linked homolog antibody
- ATP dependent helicase ATRX antibody
- ATP-dependent helicase ATRX antibody
see all
Images
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling ATRX with ab188027 at 1/200 dilution. Heat mediated antigen retrieval performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling ATRX with ab188027 showing clear nuclear (without nucleoli) staining in green. Microtubule are visualized in red.
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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 kDaDownregulation of antibody signal confirms target specificity. Remaining % intensity, relative control lane, is indicated. Anti-GAPDH monoclonal antibody was used as loading control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fallopian tube tissue labelling ATRX with ab188027 at 1/200 dilution. Heat mediated antigen retrieval performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling ATRX with ab188027 at 1/200 dilution. Heat mediated antigen retrieval performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling ATRX with ab188027 at 1/200 dilution. Heat mediated antigen retrieval performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ATRX with ab188027 showing clear nuclear (without nucleoli) staining in green. Microtubule are visualized in red.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab188027 has been referenced in 3 publications.
- Vitkunaite A et al. Intranuclear birefringent inclusions in paraffin sections by polychromatic polarization microscopy. Sci Rep 11:6275 (2021). PubMed: 33737593
- Viswanath P et al. Non-invasive assessment of telomere maintenance mechanisms in brain tumors. Nat Commun 12:92 (2021). PubMed: 33397920
- Cai HQ et al. Phosphorylated Hsp27 is mutually exclusive with ATRX loss and the IDH1R132H mutation and may predict better prognosis among glioblastomas without the IDH1 mutation and ATRX loss. J Clin Pathol 71:702-707 (2018). PubMed: 29550762