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Western blot analysis of ATR was performed with 10 µg of HeLa cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against ATR (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (NP0322BOX), XCell SureLock™ Electrophoresis System (EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (37539) for 1 hour at room temperature. ATR was detected at ~ 305 kDa using ATR Rabbit polyclonal antibody (Product # PA1-450) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Rabbit (product # 31461) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to GAPDH (36 kDa). ATR Antibody (Product # PA1-450) confirms silencing of ATR expression.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Fusion protein containing residues 400-460 of the human ATR protein.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||Assay dependent|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* 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.
PA1-450 detects ATR (Ataxia Telangiectasia Mutated (ATM) and Rad3-related protein) from human cells.
PA1-450 has been successfully used in Western blot immunofluoresence and immunoprecipitation procedures. By Western blot, this antibody detects an ~305 kDa protein representing ATR in lysate from UV irradiated K562 cells.
PA1-450 antigen is a fusion protein corresponding to amino acid residues 400-460 from human ATR.
Ataxia Telangiectasia Mutated (ATM) and Rad3-related protein (ATR) is a phosphatidylinositol kinase (PK)-related kinase which functions in response to DNA damage and repair as well as at DNA replication checkpoints during the cell cycle. ATR is a member of the DNA-PK kinases closely related to ATM and DNA-PK for which DNA stimulates the observed kinase activity. Chromosomal remodeling proteins have also been reported to associate with ATR complexes. Several known components of the NuRD complex including histone deacetylase 1 (HDAC1), HDAC2, and CHD4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Role for casein kinase 1 in the phosphorylation of Claspin on critical residues necessary for the activation of Chk1.
PA1-450 was used in western blot to study the role of casein kinase 1 in checkpoint responses to stalled DNA replication forks
|Meng Z,Capalbo L,Glover DM,Dunphy WG||Molecular biology of the cell (22:2834)||2011|
DNA-damage response, survival and differentiation in vitro of a human neural stem cell line in relation to ATM expression.
PA1-450 was used in western blot to evaluate an immortalized human neural stem-cell line as an in vitro model for the study of neuronal DNA-damage response
|Carlessi L,De Filippis L,Lecis D,Vescovi A,Delia D||Cell death and differentiation (16:795)||2009|
BRCA1 activates a G2-M cell cycle checkpoint following 6-thioguanine-induced DNA mismatch damage.
PA1-450 was used in western blot to investigate the role of BRCA1 in the G (2)-M checkpoint response to 6-thioguanine-induced DNA mismatch damage
|Yamane K,Schupp JE,Kinsella TJ||Cancer research (67:6286)||2007|
The DNA damage machinery and homologous recombination pathway act consecutively to protect human telomeres.
PA1-450 was used in western blot to investigate the involvement of DNA damage machinery and homologous recombination pathway in the maintenance of telomere integrity
|Verdun RE,Karlseder J||Cell (127:709)||2006|
Interaction between human MCM7 and Rad17 proteins is required for replication checkpoint signaling.
PA1-450 was used in western blot to study the role of hMCM7 in the DNA damage signal transmission of from active replication forks to the S-phase checkpoint.
|Tsao CC,Geisen C,Abraham RT||The EMBO journal (23:4660)||2004|
DNA replication defects, spontaneous DNA damage, and ATM-dependent checkpoint activation in replication protein A-deficient cells.
PA1-450 was used in western blot to investigate the role of replication protein A during the DNA replication.
|Dodson GE,Shi Y,Tibbetts RS||The Journal of biological chemistry (279:34010)||2004|
Determination of the catalytic activities of mTOR and other members of the phosphoinositide-3-kinase-related kinase family.
PA1-450 was used in western blot to investigate the function of proteins from phosphoinositide-3-kinase-related family
|Chiang GG,Abraham RT||Methods in molecular biology (Clifton, N.J.) (281:125)||2004|
Mismatch repair-mediated G2/M arrest by 6-thioguanine involves the ATR-Chk1 pathway.
PA1-450 was used in western blot to study the involvement of ATR-Chk1 pathway in the inhibitory effect of 6-thioguanine on DNA mismatch repair
|Yamane K,Taylor K,Kinsella TJ||Biochemical and biophysical research communications (318:297)||2004|
The ATR-p53 pathway is suppressed in noncycling normal and malignant lymphocytes.
PA1-450 was used in western blot to study the ATR-p53 pathway in normal and malignant lymphocytes
|Jones GG,Reaper PM,Pettitt AR,Sherrington PD||Oncogene (23:1911)||2004|
Quaternary structure of ATR and effects of ATRIP and replication protein A on its DNA binding and kinase activities.
PA1-450 was used in western blot to study ATR structure and its assocation with DNA.
|Unsal-Kaçmaz K,Sancar A||Molecular and cellular biology (24:1292)||2004|
An ATR- and Chk1-dependent S checkpoint inhibits replicon initiation following UVC-induced DNA damage.
PA1-450 was used in western blot to study the roles of checkpoint proteins on replication initiation after UV irradiation
|Heffernan TP,Simpson DA,Frank AR,Heinloth AN,Paules RS,Cordeiro-Stone M,Kaufmann WK||Molecular and cellular biology (22:8552)||2002|
ATR is a caffeine-sensitive, DNA-activated protein kinase with a substrate specificity distinct from DNA-PK.
PA1-450 was used in western blot to study the caffeine-sensitivity and the relation to DNA-PK of DNA-activated protein kinase ATR
|Hall-Jackson CA,Cross DA,Morrice N,Smythe C||Oncogene (18:6707)||1999|
ATR-dependent phosphorylation of FANCA on serine 1449 after DNA damage is important for FA pathway function.
PA1-450 was used in immunoprecipitation to study the role of FANCA phosphorylation in Fanconi anemia.
|Collins NB,Wilson JB,Bush T,Thomashevski A,Roberts KJ,Jones NJ,Kupfer GM||Blood (113:2181)||2009|
ATR functions as a gene dosage-dependent tumor suppressor on a mismatch repair-deficient background.
PA1-450 was used in immunoprecipitation to study the interaction between MMR proteins and ATR during the cellular response to genotoxic stress.
|Fang Y,Tsao CC,Goodman BK,Furumai R,Tirado CA,Abraham RT,Wang XF||The EMBO journal (23:3164)||2004|
Ataxia telangiectasia and Rad3-related protein; Ataxia Telangiectasia Mutated (ATM) and Rad3-Related Protein; FRAP-related protein 1; FRAP-related protein-1; MEC1, mitosis entry checkpoint 1, homolog; Serine/threonine-protein kinase ATR
ATR; FCTCS; FRP1; MEC1; SCKL; SCKL1