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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | ICC, IF, IP, WB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-740-25UG |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-phospho-ATM (Ser1981) clone 10H11.E12 (mouse monoclonal) |
| Alternate Names |
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| Background Information | Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are related kinases that regulate cell cycle checkpoints and DNA repair. Mutation in the ATM gene results in the autosomal recessive disease ataxia telangiectasia (AT). The identified substrates for ATM are p53, p95/NBS1, MDM2, Chk2, BRCA1, CtIP, 4E-BP1 and Chk1. The essential requirement for the substrates of ATM/ATR is S/TQ. Hydrophobic amino acids at positions -3 and -1, and negatively charged amino acids at position +1 are positive determinants for substrate recognition by these kinases. Positively charged residues surrounding the S/TQ are negative determinants for substrate phosphorylation. The complex phenotype of cells derived from patients with AT suggests that ATM has additional cellular substrates. In unirradiated cells, ATM is present as an inactive homodimer or multimer. Double-stranded breaks in DNA caused by ionizing radiation cause rapid ATM kinase activation through dissociation of this complex and ATM autophosphorylation at Ser1981. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Protein G purified mouse IgG in 0.1M Tris-glycine, pH 7.4, 0.15M NaCl with 0.05% sodium azide before the addition of glycerol to 30% |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Use Anti-phospho-ATM (Ser1981) Antibody, clone 10H11.E12 (Mouse Monoclonal Antibody) validated in ICC, IF, IP, WB to detect phospho-ATM (Ser1981) also known as A-T mutated. |
| Key Applications |
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| Application Notes | Immunoprecipitation: Phosphorylated ATM was immunoprecipitated from irradiated HeLa cells (Figure A, lanes 3 and 4). Immunocytochemistry: Foci are detected in irradiated human and mouse fibroblasts. Determined by an independent laboratory. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated, synthetic peptide corresponding to amino acids 1974-1988 (SLAFEEG[pS]QSTTISS) of human ATM. The immunizing sequence has 11/12 identical amino acids in mouse and rat. |
| Clone | 10H11.E12 |
| Concentration | Please refer to lot specific datasheet. |
| Host | Mouse |
| Specificity | This antibody recognizes ATM, Mr ~370 kDa. A non-specific protein was also detected, Mr ~ >400 kDa. |
| Isotype | IgG1κ |
| Species Reactivity |
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| Species Reactivity Note | Human and mouse. Predicted to cross-react with rat based on sequence homology |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene belongs to the PI3/PI4-kinase family. This protein is an important cell cycle checkpoint kinase that phosphorylates; thus, it functions as a regulator of a wide variety of downstream proteins, including tumor suppressor proteins p53 and BRCA1, checkpoint kinase CHK2, checkpoint proteins RAD17 and RAD9, and DNA repair protein NBS1. This protein and the closely related kinase ATR are thought to be master controllers of cell cycle checkpoint signaling pathways that are required for cell response to DNA damage and for genome stability. Mutations in this gene are associated with ataxia telangiectasia, an autosomal recessive disorder. Two transcript variants encoding different isoforms have been found for this gene. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: Q13315 # Serine/threonine protein kinase which activates checkpoint signaling upon double strand breaks (DSBs), apoptosis and genotoxic stresses such as ionizing ultraviolet A light (UVA), thereby acting as a DNA damage sensor. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates Ser-139 of histone variant H2AX/H2AFX at double strand breaks (DSBs), thereby regulating DNA damage response mechanism. Also involved in signal transduction and cell cycle control. May function as a tumor suppressor. Necessary for activation of ABL1 and SAPK. Phosphorylates p53/TP53, FANCD2, NFKBIA, BRCA1, CTIP, nibrin (NBN), TERF1, RAD9 and DCLRE1C. May play a role in vesicle and/or protein transport. Could play a role in T-cell development, gonad and neurological function. SIZE: 3056 amino acids; 350644 Da SUBUNIT: Exists in monomeric and tetrameric state. Binds DNA ends, p53/TP53, ABL1, BRCA1, NBN/nibrin and TERF1. Part of the BRCA1- associated genome surveillance complex (BASC), which contains BRCA1, MSH2, MSH6, MLH1, ATM, BLM, PMS2 and the RAD50-MRE11-NBN protein complex. This association could be a dynamic process changing throughout the cell cycle and within subnuclear domains. DNA damage promotes association with RAD17. Interacts with EEF1E1. This interaction, which takes place independently of TP53, is induced by DNA damage that may occur during genotoxic stress or cell growth. Interacts with DCLRE1C. Interacts with MYST1. Interacts with HTATIP. SUBCELLULAR LOCATION: Nucleus. Cytoplasmic vesicle. Note=Primarily nuclear. Found also in endocytic vesicles in association with beta-adaptin. TISSUE SPECIFICITY: Found in pancreas, kidney, skeletal muscle, liver, lung, placenta, brain, heart, spleen, thymus, testis, ovary, small intestine, colon and leukocytes.DOMAIN:SwissProt: Q13315 The FATC domain is required for interaction with HTATIP. PTM: Phosphorylated by ARK5. Autophosphorylated on Ser-1981 upon DNA damage. & Acetylated by HTATIP upon DNA damage; which is required for autophosphorylation and subsequent activation. DISEASE: SwissProt: Q13315 # Defects in ATM are the cause of ataxia telangiectasia (AT) [MIM:208900]; also known as Louis-Bar syndrome, which includes four complementation groups: A, C, D and E. This rare recessive disorder is characterized by progressive cerebellar ataxia, dilation of the blood vessels in the conjunctiva and eyeballs, immunodeficiency, growth retardation and sexual immaturity. AT patients have a strong predisposition to cancer; about 30% of patients develop tumors, particularly lymphomas and leukemias. Cells from affected individuals are highly sensitive to damage by ionizing radiation and resistant to inhibition of DNA synthesis following irradiation. & Defects in ATM contribute to T-cell acute lymphoblastic leukemia (TALL) and T-prolymphocytic leukemia (TPLL). TPLL is characterized by a high white blood cell count, with a predominance of prolymphocytes, marked splenomegaly, lymphadenopathy, skin lesions and serous effusion. The clinical course is highly aggressive, with poor response to chemotherapy and short survival time. TPLL occurs both in adults as a sporadic disease and in younger AT patients. & Defects in ATM contribute to B-cell non-Hodgkin lymphomas (BNHL), including mantle cell lymphoma (MCL). & Defects in ATM contribute to B-cell chronic lymphocytic leukemia (BCLL). BCLL is the commonest form of leukemia in the elderly. It is characterized by the accumulation of mature CD5+ B lymphocytes, lymphadenopathy, immunodeficiency and bone marrow failure. SIMILARITY: Belongs to the PI3/PI4-kinase family. ATM subfamily. & Contains 1 FAT domain. & Contains 1 FATC domain. & Contains 1 PI3K/PI4K domain. |
| Molecular Weight | ~370 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated by immunoblot on in crude lysates from irradiated HeLa cells. Western Blot Analysis: 0.5 µg/mL of this lot detected phosphorylated ATM in crude lysates from irradiated HeLa cells. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 25 μg |