Immunofluorescent analysis of ATRIP(green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 5% normal goat serum (Product #31873) for 15 minutes at room temperature. Cells were then probed with a rabbit polyclonal antibody recognizing ATRIP (Product# PA1-519), at a dilution of 1:400 for at least 1 hour at room temperature. Cells were then washed with PBS and incubated with DyLight 488 goat-anti-rabbit secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Immunogen corresponds to amino acids 1-182 of human ATRIP.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.02% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunocytochemistry (ICC)||See 1 publications below|
PA1-519 detects human and mouse ATRIP.
PA1-519 has been successfully used in Western blot, immunoprecipitation and immunofluorescence procedures. By Western blot, this antibody detects an ~ 86 kDa protein from human HeLa cells.
PA1-519 antigen corresponds to amino acids 1-182 of human ATRIP
ATR- interacting protein (ATRIP) is an essential component of the DNA damage checkpoint pathway. ATRIP, phosphorylated by ataxia telangiectasia mutated and Rad3 kinase (ATR), regulates ATR expression, signifying their mutual dependence. The ATR-ATRIP protein kinase complex is essential for the cellular response to replication stress and DNA damage.
It has been shown, in vitro, that replication protein A (RPA) stimulates the binding of ATRIP to single-stranded DNA. This enables the ATR-ATRIP complex to associate with DNA and stimulates phosphorylation of the RAD17 protein that is bound to DNA. ATR/ATRIP exist as a large molecular weight complex that can bind single-stranded DNA cellulose in vitro. Neither ATR nor ATRIP are able to bind DNA individually, nor do they bind DNA in a cooperative manner.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Replication stress is a potent driver of functional decline in ageing haematopoietic stem cells.
PA1-519 was used in immunocytochemistry to study why HSC function declines with age
|Flach J,Bakker ST,Mohrin M,Conroy PC,Pietras EM,Reynaud D,Alvarez S,Diolaiti ME,Ugarte F,Forsberg EC,Le Beau MM,Stohr BA,Méndez J,Morrison CG,Passegué E||Nature (512:198)||2014|