Phospho-gamma H2A.X (S139) ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-gamma H2A.X (S139) in Human in Cell Lysate samples.
Application | Reactivity | Dilution info | Notes |
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
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.
H2AFX, H2AX, Histone H2AX, H2a/x, Histone H2A.X, H2AS139p, H2AXS139p, H2A.XS139p
Phospho-gamma H2A.X (S139) ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-gamma H2A.X (S139) in Human in Cell Lysate samples.
Phospho-gamma H2A.X (S139) ELISA Kit (ab279816) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated gamma H2A.X protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-gamma H2A.X. An anti-pan gamma H2A.X antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and gamma H2A.X present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-gamma H2A.X (S139) antibody is used to detect phosphorylated gamma H2A.X. After washing away unbound antibody, HRP conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of phospho-gamma H2A.X bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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Gamma H2A.X also known as phospho H2A.X or γH2A.X is a phosphorylated form of the histone variant H2A.X. It has a molecular weight of about 14 kilodaltons and occurs primarily in they nucleus. When DNA double-strand breaks (DSBs) occur serine 139 in H2A.X undergoes rapid phosphorylation resulting in gamma H2A.X. This modification happens swiftly at the site of damage and gamma H2A.X spreads over a large chromatin area facilitating the recruitment of DNA repair proteins. Gamma H2A.X staining typically evaluated using gamma H2A.X immunofluorescence techniques aids in identifying the presence and extent of DNA damage.
Gamma H2A.X plays a role in DNA damage response and repair. It does not operate alone; it acts as part of a complex with other repair proteins. The formation of gamma H2A.X foci at DNA damage sites creates a signal attracting repair factors that help maintain genome stability. Its interaction with MDC1 and ATM proteins exemplifies its significant role in orchestrating an effective response to DNA damage. Beyond DNA repair gamma H2A.X influences cell cycle checkpoints permitting cells to pause and repair before proceeding with division.
Gamma H2A.X plays a pivotal role in the DNA damage response (DDR) pathway. This pathway is essential for detecting and repairing DNA lesions to uphold genomic integrity. Within the DDR pathway gamma H2A.X is closely associated with proteins such as NBS1 and BRCA1 which assist in repairing double-strand breaks. In addition gamma H2A.X is integral to the ATM-ATR signaling pathway where its activation promotes cell survival following genotoxic stress by signaling for damage repair or triggering apoptosis.
Gamma H2A.X has connections to cancer and neurodegeneration. Aberrant DNA repair pathways often indicated by persistent gamma H2A.X signals correlate with tumor formation and progression. For instance a failure to repair DNA damage effectively can lead to mutations that drive cancer development. Gamma H2A.X also links to neurodegenerative diseases where dysregulated DNA repair contributes to neuronal cell death. Proteins like p53 which regulate cell cycle and apoptosis further connect to gamma H2A.X bridging its role in disease pathogenesis.
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Positive Control.
Jurkat cells were treated with CPT at 37°C for 16 hours.
Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
Jurkat cells treated with/without CPT. Jurkat cells were untreated or treated with CPT for 16 hours.
Jurkat cells treated with/without CPT. Jurkat cells were untreated or treated with CPT for 16 hours.
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