Immunofluorescence analysis of Phospho-ATM pSer1981 was performed using 70% confluent log phase HeLa cells irradiated with UV for 4 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-ATM (Ser 1981) (10H11) Mouse Monoclonal Antibody (MA12020) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the untreated cells. Panel f represents the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues S(1974) L A F E E S(p) Q S T T I S S(1988) of human ATM Kinase protein.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:500 - 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.
MA1-2020 detects the phospho-ATM kinase in human and mouse samples.
MA1-2020 has been successfully used in immunofluorescence, immunoprecipitation and Western blot procedures. By Western blot this antibody detects a ~370 kDa protein representing the phospho-ATM kinase in crude lysates from gamma irradiated HeLa cells. In immunofluorescence procedures, MA1-2020 recognizes the phospho-ATM kinase in irradiated human and mouse fibroblasts.
The MA1-2020 immunogen is a phosphorylated synthetic peptide corresponding to the residues S(1974) L A F E E S(p) Q S T T I S S(1988) of human ATM Kinase protein.
Ataxia-telangiectasia (A-T) is a rare autosomal recessive disorder characterized by progressive neurologic degeneration, immunologic deficiency, and an increased risk of lymphoid cancer. The A-T mutant (ATM) gene codes for a protein belonging to the phosphoinositide 3-kinase (PI3K) superfamily. ATM phosphorylates proteins instead of lipid and has many downstream targets that act as cell-cycle regulators including: P53, Mdm2, BRCA1, and SMC1. The ATM protein is responsible for repairing double-stranded DNA breaks that occur because of ionizing radiation and other mutagens. Studies have shown that ATM becomes autophosphorylated and upregulated by exposure to ionizing radiation. ATM and quote;s role in cancer is still being elucidated.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A localized nucleolar DNA damage response facilitates recruitment of the homology-directed repair machinery independent of cell cycle stage.
MA1-2020 was used in immunocytochemistry to study the response to double-stranded breaks at the nucleolar organizer regions.
|van Sluis M,McStay B||Genes and development (29:1151)||2015|
2-Hydroxyethyl methacrylate-induced apoptosis through the ATM- and p53-dependent intrinsic mitochondrial pathway.
MA1-2020 was used in western blot to study the mechanism for 2-hydroxyethyl methacrylate-induced apoptosis
|Schweikl H,Petzel C,Bolay C,Hiller KA,Buchalla W,Krifka S||Biomaterials (35:2890)||2014|