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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | FC, ICC | M | FITC | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 16-202A |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, FITC conjugate |
| Alternate Names |
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| Background Information | Histone H2A is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N terminal tail H2A is involved with the structure of the nucleosomes of the beads on a string structure. |
| Product Information | |
|---|---|
| Format | FITC |
| Control |
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| Presentation | Purified FITC-conjugated mouse monoclonal IgG1 in buffer containing PBS with 0.05% sodium azide. Frozen solution. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Use Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, FITC conjugate (mouse monoclonal antibody) validated in FC, ICC to detect phospho-Histone H2A.X (Ser139) also known as H2AXS139P, H2AX histone. |
| Key Applications |
|
| Application Notes | Immunocytochemistry: 2-4 μg/mL of a previous lot detected phospho-histone H2A.X in etoposide-treated HeLa cells fixed with 95% ethanol/5% acetic acid (10 minutes), followed by 1% formaldehyde, 0.25% Triton® X-100 in TBS (5 min.). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated, synthetic peptide (CKATQA[PS]QEY) corresponding to amino acids 134-142 of human histone H2A.X. The immunizing sequence has 8 identical amino acids in yeast and mouse. Clone JBW301. |
| Epitope | Ser139 |
| Clone | clone JBW301 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | Recognizes Histone H2A.X phosphorylated at Ser139, MW 15 kDa. |
| Isotype | IgG1 |
| Species Reactivity |
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| Species Reactivity Note | Human. Broad species cross-reactivity is expected based on conservation of sequence homology. |
| Antibody Type | Monoclonal 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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| Modifications |
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| Purification Method | Protein g Purified |
| 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. |
| Molecular Weight | 15 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Flow Cytometry to detect Log phase Jurkat cells treated with staurosporine. Flow Cytometry: Log phase Jurkat cells were treated with staurosporine (1 μg/mL) for the indicated time. Histone H2A.X (Ser 139) phosphorylation was detected as described in the manual for the H2A.X Phosphorylation Assay Kit (Flow Cytometry), Catalog # 17-344. Cells were analyzed on a Becton-Dickinson FACS-Calibur flow cytometer. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |