Anti-ATRX antibody (ab204169)
Key features and details
- Rabbit polyclonal to ATRX
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-ATRX antibody
See all ATRX primary antibodies -
Description
Rabbit polyclonal to ATRX -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Chimpanzee -
Immunogen
Recombinant fragment corresponding to Human ATRX aa 2250-2450.
Database link: P46100 -
Positive control
- WB: A549 whole cell lysates; IHC-P: Human uterine cervix, glioma, cerebral cortex tissue; ICC :A549 whole 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.
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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: 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
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 ab204169 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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ICC/IF |
Use a concentration of 0.25 - 2 µg/ml.
Fixation/Permeabilization: PFA/Triton X-100 |
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IHC-P |
1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use a concentration of 0.04 - 0.4 µg/ml. Predicted molecular weight: 282 kDa.
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Notes |
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ICC/IF
Use a concentration of 0.25 - 2 µg/ml. Fixation/Permeabilization: PFA/Triton X-100 |
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use a concentration of 0.04 - 0.4 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: 449625 Chimpanzee
- Entrez Gene: 546 Human
- Entrez Gene: 22589 Mouse
- Entrez Gene: 246284 Rat
- Omim: 301040 Human
- SwissProt: Q7YQM4 Chimpanzee
- SwissProt: P46100 Human
- SwissProt: Q61687 Mouse
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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All lanes : Anti-ATRX antibody (ab204169) at 0.4 µg/ml
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : HCT 116 cell lysate
Lane 3 : U-2 OS cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 282 kDa
Observed band size: 150,180,350 kDa why is the actual band size different from the predicted?Western blot: Anti-ATRX antibody staining at 0.4 ug/ml, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to ATRX. A band was observed at 150/180/350 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in ATRX knockout cell line ab287231. To generate this image, wild-type and ATRX knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 12 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-ATRX antibody (ab204169) at 0.4 µg/ml
Lane 1 : Control siRNA - A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : siRNA probe #1 - A549 whole cell lysate
Lane 3 : siRNA probe #2 - A549 whole cell lysate
Predicted band size: 282 kDaLoading control: Anti-GAPDH.
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Immunohistochemical analysis of human uterine cervix tissue labeling ATRX in the nucleus of squamous epithelial cells with ab204169 at a 1/500 dilution.
Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemical analysis of A549 (Human lung carcinoma cell line) whole cells labeling ATRX in the nucleoplasm and nuclear bodies with ab204169 at 2 µg/ml.
Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of human glioma tissue labeling ATRX in the nucleus of tumor cells with ab204169 at a 1/500 dilution.
Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of human cerebral cortex tissue labeling ATRX in the nucleus of neurons and glial cells with ab204169 at a 1/500 dilution.
Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of human liver tissue labeling ATRX in the hepatocytes with ab204169 at a 1/500 dilution. No positivity as expected.
Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab204169 has been referenced in 1 publication.
- Ban X et al. Alternative Lengthening of Telomeres Phenotype Predicts Progression Risk in Noninsulinomas in a Chinese Cohort. Neuroendocrinology 112:510-522 (2022). PubMed: 34348341