Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y170] to ATM
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Reacts with: Human
Product nameAnti-ATM antibody [Y170]
See all ATM primary antibodies
DescriptionRabbit monoclonal [Y170] to ATM
SpecificityThis antibody is specific for ATM.
Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
Unsuitable for: IP
Species reactivityReacts with: Human
- WB: 293 cell lysate. Flow Cyt: HeLa cells ICC/IF: HeLa and HepG2 cells
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab32420 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 350 kDa.Can be blocked with Human ATM peptide (ab170988).|
|IHC-P||1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
|ICC/IF||1/250 - 1/500.
FunctionSerine/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 plays a role in pre-B cell allelic exclusion, a process leading to expression of a single immunoglobulin heavy chain allele to enforce clonality and monospecific recognition by the B-cell antigen receptor (BCR) expressed on individual B lymphocytes. After the introduction of DNA breaks by the RAG complex on one immunoglobulin allele, acts by mediating a repositioning of the second allele to pericentromeric heterochromatin, preventing accessibility to the RAG complex and recombination of the second allele. 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. Plays a role in replication-dependent histone mRNA degradation. Binds DNA ends.
Tissue specificityFound in pancreas, kidney, skeletal muscle, liver, lung, placenta, brain, heart, spleen, thymus, testis, ovary, small intestine, colon and leukocytes.
Involvement in diseaseDefects 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.
Note=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.
Note=Defects in ATM contribute to B-cell non-Hodgkin lymphomas (BNHL), including mantle cell lymphoma (MCL).
Note=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.
Sequence similaritiesBelongs to the PI3/PI4-kinase family. ATM subfamily.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 1 PI3K/PI4K domain.
DomainThe FATC domain is required for interaction with KAT5.
modificationsPhosphorylated by NUAK1/ARK5. Autophosphorylation on Ser-367, Ser-1893, Ser-1981 correlates with DNA damage-mediated activation of the kinase.
Acetylation, on DNA damage, is required for activation of the kinase activity, dimer-monomer transition, and subsequent autophosphorylation on Ser-1981. Acetylated in vitro by KAT5/TIP60.
Cellular localizationNucleus. Cytoplasmic vesicle. Primarily nuclear. Found also in endocytic vesicles in association with beta-adaptin.
- Information by UniProt
- A-T mutated antibody
- A-T mutated homolog antibody
- AT mutated antibody
Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling ATM with ab32420 at 1/500. Goat anti rabbit IgG(Alexa Fluor® 488), ab150077 at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nulei were counterstained blue with DAPI.
All lanes : Anti-ATM antibody [Y170] (ab32420) at 1/3000 dilution (Purified)
Lane 1 : HeLa cell lysate
Lane 2 : HEK293 cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 350 kDa
Observed band size: 370 kDa why is the actual band size different from the predicted?
Immunocytochemistry/ Immunofluorescence analysis of HepG2 cells labeling ATM with ab32420 at 1/500. Goat anti rabbit IgG(Alexa Fluor® 488), ab150077 at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nulei were counterstained blue with DAPI.
DNA repair proteins accumulate at MVM APAR bodies
Repair proteins accumulate at APAR bodies. NB324K cells were infected with MVMp (MOI of 10) for 16 hr before being fixed and processed for immunofluorescence. Cells were stained with the indicated antibodies to mark DDR repair proteins. APAR bodies were detected with antibodies to NS1. Nuclei were stained with DAPI. All images were captured using an objective of 63×.
Cells were fixed with 4% paraformaldehyde for 15 minutes and permeabilized with 0.5% Triton X-100 in PBS for 15 minutes.
(Image shows the right-hand panel of Figure 2A)
Formaldehyde-fixed human colon tissue stained for ATM using ab32420 at 1/100 dilution in immunohistochemical analysis.
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling ATM with purified ab32420 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ATM with ab32420 at 1:100 dilution. Tissue underwent antigen retrieval using Tris/EDTA Buffer (pH9.0). The section was counterstained with haematoxylin.
Formaldehyde-fixed human serous ovarian tumor tissue stained for ATM using ab32420 at 1/50 dilution in immunohistochemical analysis.
Immunohistochemical analysis of paraffin-embedded Human normal breast tissue labeling ATM with ab32420 at 1:100 dilution. Tissue underwent antigen retrieval using Tris/EDTA Buffer (pH9.0). The section was counterstained with haematoxylin.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32420 has been referenced in 80 publications.
- Lowe DJ et al. Chronic irradiation of human cells reduces histone levels and deregulates gene expression. Sci Rep 10:2200 (2020). PubMed: 32042076
- Cao Y et al. CENPO expression regulates gastric cancer cell proliferation and is associated with poor patient prognosis. Mol Med Rep 20:3661-3670 (2019). PubMed: 31485675
- Rehman A et al. The desmosomal cadherin desmoglein-3 acts as a keratinocyte anti-stress protein via suppression of p53. Cell Death Dis 10:750 (2019). PubMed: 31582719
- Ali R et al. ATM Regulated PTEN Degradation Is XIAP E3 Ubiquitin Ligase Mediated in p85a Deficient Cancer Cells and Influence Platinum Sensitivity. Cells 8:N/A (2019). PubMed: 31635307
- Angell HK et al. PD-L1 and immune infiltrates are differentially expressed in distinct subgroups of gastric cancer. Oncoimmunology 8:e1544442 (2019). PubMed: 30729066