Phospho-ATR (T1989) and Total ATR ELISA Kit (ab279742)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
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Product name
Phospho-ATR (T1989) and Total ATR ELISA Kit
See all ATR kits -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
Phospho-ATR (T1989) and Total ATR ELISA Kit (ab279742) 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 ATR 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-ATR and total ATR. An anti-pan ATR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and ATR present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-ATR (T1989) antibody is added to detect phosphorylated ATR. In the remaining wells, mouse anti-pan-ATR antibody is used to detect pan ATR. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-mouse 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 ATR (T1989) or pan ATR 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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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 20X Wash Buffer Concentrate 1 x 25ml 2X Cell lysate buffer 1 x 10ml 5X Assay Diluent 1 x 15ml HRP-conjugated anti-mouse IgG concentrate (1000X) 1 vial HRP-conjugated anti-rabbit IgG concentrate (500X) 1 vial Mouse anti-pan-ATR detection antibody 1 vial Pan-ATR Coated Microplate 1 unit Positive Control - T47D cell lysate 1 vial Rabbit anti-phospho-ATR (T1989)-antibody 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
Function
Serine/threonine protein kinase which activates checkpoint signaling upon genotoxic stresses such as ionizing radiation (IR), ultraviolet light (UV), or DNA replication stalling, thereby acting as a DNA damage sensor. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates BRCA1, CHEK1, MCM2, RAD17, RPA2, SMC1 and p53/TP53, which collectively inhibit DNA replication and mitosis and promote DNA repair, recombination and apoptosis. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX at sites of DNA damage, thereby regulating DNA damage response mechanism. Required for FANCD2 ubiquitination. Critical for maintenance of fragile site stability and efficient regulation of centrosome duplication. -
Tissue specificity
Ubiquitous, with highest expression in testis. Isoform 2 is found in pancreas, placenta and liver but not in heart, testis and ovary. -
Involvement in disease
Defects in ATR are a cause of Seckel syndrome type 1 (SCKL1) [MIM:210600]. SCKL1 is a rare autosomal recessive disorder characterized by growth retardation, microcephaly with mental retardation, and a characteristic 'bird-headed' facial appearance. -
Sequence similarities
Belongs to the PI3/PI4-kinase family. ATM subfamily.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 2 HEAT repeats.
Contains 1 PI3K/PI4K domain. -
Post-translational
modificationsPhosphorylated; autophosphorylates in vitro. -
Cellular localization
Nucleus. Nucleus > PML body. Depending on the cell type, it can also be found in PML nuclear bodies. Recruited to chromatin during S-phase. Redistributes to discrete nuclear foci upon DNA damage, hypoxia or replication fork stalling. - Information by UniProt
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Alternative names
- Ataxia telangiectasia and Rad3 related
- Ataxia telangiectasia and Rad3-related protein
- ATR
see all -
Database links
- Entrez Gene: 545 Human
- Omim: 601215 Human
- SwissProt: Q13535 Human
- Unigene: 271791 Human
Images
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T47D cells were exposed to 50 J/m2 of UV light followed by a 4 hours recovery period.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
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T47D cells were untreated or treated with UV.
Cell lysates were analyzed using this phosphoELISA and Western Blot.