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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | PIA, WB | Rb | Serum | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-627 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-Histone H2A.X Antibody |
| Alternate Names |
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| Background Information | Histone H2AX (UniProt: P16104; also known as H2a/x, Histone H2A.X) is encoded by the H2AFX (also known as H2AX) gene (Gene ID: 3014) in human. Histones are highly conserved basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. They 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. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which DNA is wrapped in repeating units, called nucleosomes, which limits DNA accessibility to the cellular machineries that require DNA as a template. The histone H2A.X is a variant member of the H2A family of histones that is distinguished from other H2A histones by a unique carboxy-terminal sequence. This C-terminal tail becomes phosphorylated following double-stranded DNA breaks following irradiation or apoptosis. This unique sequence is highly conserved throughout eukaryotic evolution and is rapidly phosphorylated by ATM or ATR in response to DNA double-strand breaks. H2A.X phosphorylation is important in the formation of a stable repair complex at the site of DNA damage. |
| Product Information | |
|---|---|
| Format | Serum |
| Control |
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| Presentation | Rabbit antiserum containing 0.05% sodium azide and 30% glycerol. Liquid at -20°C. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Histone H2A.X Antibody, Cat. No. 07-627, is a highly specific rabbit polyclonal antibody that targets Histone H2A.X and has been tested for use in Immunohistochemistry (Paraffin) and Western Blotting. |
| Key Applications |
|
| Application Notes | Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected Histone H2A.X in human pancreas and human colon tissue sections. Western Blotting Analysis: A 1:500 dilution from a representative lot detected Histone H2A.X in H2A and H2A.X recombinant protein. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated, synthetic peptide (CSATVGPKAPSGGKKA) corresponding to amino acids 121-135 of human histone H2A.X, with an N-terminal cysteine added for conjugation purposes. The immunizing sequence has 13/15 identical amino acids in mouse. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody detects Histone H2AX in human cells. It targets an epitope with in 15 amino acids from the C-terminal region. |
| Isotype | IgG |
| Species Reactivity |
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| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | 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. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. |
| Gene Symbol |
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| Purification Method | Unpurified |
| UniProt Number |
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| UniProt Summary | FUNCTION:SwissProt: P16104 # Variant histone H2A which replaces conventional H2A in a subset of 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. 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. SIZE: 143 amino acids; 15145 Da SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with numerous proteins required for DNA damage signaling and repair when phosphorylated on Ser-140. These include MDC1, TP53BP1, BRCA1 and the MRN complex, composed of MRE11A, RAD50, and NBN. Interaction with the MRN complex is mediated at least in part by NBN. Also interacts with DHX9/NDHII when phosphorylated on Ser-140. SUBCELLULAR LOCATION: Nucleus.DEVELOPMENTAL STAGE: Synthesized in G1 as well as in S-phase. DOMAIN: SwissProt: P16104 The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family. PTM: Phosphorylated on Ser-140 (to form gamma-H2AFX) 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. Phosphorylation of Ser-140 in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation may also be required for transcriptional repression of unsynapsed chromatin and meiotic sex chromosome inactivation (MSCI), whereby the X and Y chromosomes condense in pachytene to form the heterochromatic XY-body. During immunoglobulin class switch recombination in lymphocytes, Ser-140 phosphorylation may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. & Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression (By similarity). SIMILARITY: Belongs to the histone H2A family. ... hide » see more » Entrez Gene Number: NP_002096 Entrez Gene Summary 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. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. |
| Molecular Weight | ~15 kDa; 15.15 kDa calculated. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in Jurkat cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Histone H2A.X in Jurkat cell lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 200 µL |