Phospho-ATM (S1981) and Total ATM ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-ATM (S1981) and Total ATM in Human in Cell Lysate samples.
Colorimetric
Cell Lysate
Semi-quantitative
Human
Application | Reactivity | Dilution info | Notes |
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Phospho-ATM (S1981) and Total ATM ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-ATM (S1981) and Total ATM in Human in Cell Lysate samples.
Colorimetric
Cell Lysate
Semi-quantitative
Human
Pre-coated microplate (12 x 8 well strips)
Blue Ice
-20°C
-20°C
-20°C
Phospho-ATM (S1981) and Total ATM ELISA Kit (ab279740) 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 ATM 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-ATM and total ATM. An anti-pan ATM antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and ATM present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho ATM (S1981) antibody is added to detect phosphorylated ATM (S1981). In the remaining wells, biotinylated anti-pan-ATM antibody is used to detect pan ATM. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin 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 ATM (S1981) or pan ATM 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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ATM also known as Ataxia Telangiectasia Mutated is a protein kinase with a molecular weight of approximately 370 kDa. ATM protein primarily resides in the cell nucleus and functions as a critical regulator of the cell cycle. It plays a significant role in the detection of DNA damage and initiation of repair processes. As part of its mechanical functions ATM phosphorylates serine and threonine residues on various substrates most notably in response to double-strand breaks in DNA. This activity is important for maintaining genomic stability.
ATM acts as a coordinator in cellular response to DNA damage highly interacting with multiple components of the DNA repair machinery. It forms a complex with proteins like NBS1 and MRN complex facilitating repair by recruiting and activating other proteins involved in homologous recombination and non-homologous end joining pathways. ATM also modulates p53 activity a primary response factor in cellular stress management linking ATM to control of cell cycle arrest and apoptosis. This positions ATM as an integral part of maintaining cellular integrity in face of genomic insult.
ATM integrates neatly within the DNA damage response and cell cycle control pathways. ATM's operative relationship with the MRN complex and its role in the PI3K-related protein kinase family helps initiate appropriate repair processes upon DNA damage detection. Additionally ATM regulates the activity of proteins such as Chk2 which further propagates signals to p53 influencing decisions between cell cycle arrest and apoptosis. These interactions link ATM closely to essential processes like DNA repair and cell survival highlighting its role in genomic maintenance.
ATM mutations or dysregulation leads to Ataxia Telangiectasia an autosomal recessive disorder characterized by neurodegeneration immune deficiencies and cancer predisposition. ATM dysfunction also connects to cancer development particularly breast cancer where it transmits signals involving BRCA1 contributing to DNA repair through homologous recombination. Understanding ATM dynamics and related pathways has important implications for developing therapeutic strategies to manage or mitigate effects associated with its dysfunction.
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Positive Control.
T47D cells were treated with UV.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
UV treated T47D cells treated with/without LPP.
T47D cells were treated with UV.
Cell lysates were untreated or treated with Lambda Protein Phosphatase (LPP) and analyzed using this phosphoELISA and Western Blot.
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