Immunofluorescence (IF)
(Histone H2AX phospho Ser139 antibody tested by immunofluorescence. HeLa cells stained with Histone H2AX phospho Ser139 antibody (1:500 dilution) using MAX Stain Immunofluorescence Tools. The HeLa cells were blocked with MAXblock Blocking Medium and stained with Histone H2AX phospho Ser139 antibody. Panel A: Untreated HeLa cells. Panel B: Cells fixed and stained 90 minutes after 3 Gy ionizing radiation treatment. Top images: Cells were stained with DAPI. Middle images: Same cells stained with Histone H2AX phospho Ser139 antibody. Bottom images: Merge of both images above. Images were made using Zeiss Axiovision with equivalent acquisition settings for direct comparison. Note Panel B-middle image, which shows intense nuclear clustering of ionizing radiation-induced phosphorylation of Ser139 of H2AX. In contrast, Panel A-middle image shows no detectable phosphorylated H2AX.)
Rabbit Histone H2A.XS139ph (H2AX) Polyclonal Antibody | anti-H2A.XS139ph antibody
Histone H2A.XS139ph antibody (pAb)(H2AX)
ICC/IF: 1:500 dilution
Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western Blot.
Immunofluorescence (IF)
(Histone H2AX phospho Ser139 antibody tested by immunofluorescence. HeLa cells stained with Histone H2AX phospho Ser139 antibody (1:500 dilution) using MAX Stain Immunofluorescence Tools. The HeLa cells were blocked with MAXblock Blocking Medium and stained with Histone H2AX phospho Ser139 antibody. Panel A: Untreated HeLa cells. Panel B: Cells fixed and stained 90 minutes after 3 Gy ionizing radiation treatment. Top images: Cells were stained with DAPI. Middle images: Same cells stained with Histone H2AX phospho Ser139 antibody. Bottom images: Merge of both images above. Images were made using Zeiss Axiovision with equivalent acquisition settings for direct comparison. Note Panel B-middle image, which shows intense nuclear clustering of ionizing radiation-induced phosphorylation of Ser139 of H2AX. In contrast, Panel A-middle image shows no detectable phosphorylated H2AX.)
Immunofluorescence (IF)
(Histone H2AX phospho Ser139 antibody tested by immunofluorescence. HeLa cells stained with Histone H2AX phospho Ser139 antibody (1:500 dilution) using MAX Stain Immunofluorescence Tools. The HeLa cells were blocked with MAXblock Blocking Medium and stained with Histone H2AX phospho Ser139 antibody. Panel A: Untreated HeLa cells. Panel B: Cells fixed and stained 90 minutes after 3 Gy ionizing radiation treatment. Top images: Cells were stained with DAPI. Middle images: Same cells stained with Histone H2AX phospho Ser139 antibody. Bottom images: Merge of both images above. Images were made using Zeiss Axiovision with equivalent acquisition settings for direct comparison. Note Panel B-middle image, which shows intense nuclear clustering of ionizing radiation-induced phosphorylation of Ser139 of H2AX. In contrast, Panel A-middle image shows no detectable phosphorylated H2AX.)
Immunofluorescence (IF)
(Histone H2AX phospho Ser139 antibody tested by immunofluorescence. Etoposide-treated HeLa cells stained with (1:500 dilution, red) and counterstained with DAPI (blue).)
Immunofluorescence (IF)
(Histone H2AX phospho Ser139 antibody tested by immunofluorescence. Etoposide-treated HeLa cells stained with (1:500 dilution, red) and counterstained with DAPI (blue).)
Western Blot (WB)
(Histone H2AX phospho Ser139 antibody tested by Western blot. Western blot: Nuclear extract of U2OS cells (20 ug per lane) probed with Histone H2AX phospho Ser139 polyclonal antibody (1:500 dilution). Lane 1: untreated cells Lane 2: cells treated with camptothecin)
Western Blot (WB)
(Histone H2AX phospho Ser139 antibody tested by Western blot. Western blot: Nuclear extract of U2OS cells (20 ug per lane) probed with Histone H2AX phospho Ser139 polyclonal antibody (1:500 dilution). Lane 1: untreated cells Lane 2: cells treated with camptothecin)
Background: Histone H2AX phospho Ser139 (H2AX, H2A histone family member X) replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries that require DNA as a template. Histones thereby play a central role in transcriptional regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called the histone code, and nucleosome remodeling. Histone H2AX is required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation, and for efficient repair of DNA double-strand breaks (DSBs), specifically when modified by C-terminal phosphorylation.
NCBI and Uniprot Product Information
Uniprot Description
H2AX: a histone that replaces conventional H2A in a subset of nucleosomes. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Phosphorylated on S139 by ATM and DNA-PK in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Dephosphorylation of S139 by PP2A is required for DNA DSB repair. Apparently phosphorylated on Y143 by WSTF, determining the relative recruitment of either DNA repair or pro-apoptotic factors. H2AXpY142 favors the recruitment of pro-apoptotic factors APBB1 and JNK1. In contrast, dephosphorylation of pY143 by EYA phosphatases favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated pS139. Monoubiquitination of K119 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'K63' linkages by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage.
Protein type: Helicase; DNA-binding; DNA repair, damage
Chromosomal Location of Human Ortholog: 11q23.3
Cellular Component: male germ cell nucleus; nucleoplasm; XY body; chromosome, telomeric region; condensed nuclear chromosome; nuclear chromatin; replication fork; nucleosome; nucleus
Molecular Function: protein binding; enzyme binding; DNA binding; histone binding; protein heterodimerization activity; damaged DNA binding
Biological Process: nucleosome assembly; positive regulation of DNA repair; meiotic cell cycle; double-strand break repair; DNA damage checkpoint; spermatogenesis; response to ionizing radiation; DNA repair; response to DNA damage stimulus; double-strand break repair via homologous recombination
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Product Notes
The H2A.XS139ph h2afx (Catalog #AAA388374) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Histone H2A.XS139ph antibody (pAb)(H2AX) reacts with Human, Wide Range Predicted and may cross-react with other species as described in the data sheet. AAA Biotech's Histone H2A.XS139ph (H2AX) can be used in a range of immunoassay formats including, but not limited to, Dot Blot (DB), Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blot (WB). Applications Validated: ICC/IF: 1:500 dilution Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western Blot. Researchers should empirically determine the suitability of the H2A.XS139ph h2afx for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Histone H2A.XS139ph (H2AX), Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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