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Anti-ATM (phospho S1981)抗体[10H11.E12] (ab36810)

Reviews (2)Q&A (11)References (75)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
  • Western blot - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
  • Flow Cytometry (Intracellular) - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
  • Western blot - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)

Key features and details

  • Mouse monoclonal [10H11.E12] to ATM (phospho S1981)
  • Suitable for: WB, Flow Cyt (Intra), IHC-P
  • Reacts with: Human
  • Isotype: IgG1

概述

  • 产品名称

    Anti-ATM (phospho S1981)抗体[10H11.E12]
    参阅全部 ATM 一抗

  • 描述

    小鼠单克隆抗体[10H11.E12] to ATM (phospho S1981)

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, Flow Cyt (Intra), IHC-Pmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide corresponding to Human ATM.
    Database link: Q13315

  • 阳性对照

    • Irradiated normal human fibroblasts (no reactivity against non-irradiated cell extracts).

  • 常规说明

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab36810于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/1000. Detects a band of approximately 370 kDa (predicted molecular weight: 370 kDa).

Abcam recommends using 3% Milk as the blocking agent.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P (1)
Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
说明
WB
1/1000. Detects a band of approximately 370 kDa (predicted molecular weight: 370 kDa).

Abcam recommends using 3% Milk as the blocking agent.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能

    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 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.

  • 组织特异性

    Found in pancreas, kidney, skeletal muscle, liver, lung, placenta, brain, heart, spleen, thymus, testis, ovary, small intestine, colon and leukocytes.

  • 疾病相关

    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.
    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.

  • 序列相似性

    Belongs to the PI3/PI4-kinase family. ATM subfamily.
    Contains 1 FAT domain.
    Contains 1 FATC domain.
    Contains 1 PI3K/PI4K domain.

  • 结构域

    The FATC domain is required for interaction with KAT5.

  • 翻译后修饰

    Phosphorylated 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.

  • 细胞定位

    Nucleus. Cytoplasmic vesicle. Primarily nuclear. Found also in endocytic vesicles in association with beta-adaptin.
  • Information by UniProt

  • 数据库链接

    • 别名

      • A-T mutated antibody
      • A-T mutated homolog antibody
      • AT mutated antibody
      • AT1 antibody
      • ATA antibody
      • Ataxia telangiectasia mutated antibody
      • Ataxia telangiectasia mutated gene antibody
      • Ataxia telangiectasia mutated homolog (human) antibody
      • Ataxia telangiectasia mutated homolog antibody
      • ATC antibody
      • ATD antibody
      • ATDC antibody
      • ATE antibody
      • ATM antibody
      • ATM serine/threonine kinase antibody
      • ATM_HUMAN antibody
      • DKFZp781A0353 antibody
      • MGC74674 antibody
      • OTTHUMP00000232981 antibody
      • Serine protein kinase ATM antibody
      • Serine-protein kinase ATM antibody
      • Serine/threonine-protein kinase ATM antibody
      • Tefu antibody
      • TEL1 antibody
      • TEL1, telomere maintenance 1, homolog antibody
      • TELO1 antibody
      • Telomere fusion protein antibody
      see all

    图片

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)

      ab36810 staining human colon tissue. Staining is localized to the nucleus.
      Left panel: with primary antibody at 4 µg/ml. Right panel: Isotype control.
      Sections were stained using an automated system DAKO Autostainer Plus at room temperature. Sections were rehydrated and antigen retrieved with the DAKO 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with hematoxylin and coverslipped under DePeX.

      Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

    • Western blot - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
      Western blot - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
      All lanes : Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810) at 10 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Extract from patient with Ataxia-Telangiectasia whole cell lysate
      Lane 3 : Irradiated HeLa Whole Cell Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Lanes 1 & 3 : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
      Lane 2 : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under non-reducing conditions.

      Predicted band size: 370 kDa
      Observed band size: 370 kDa
      Additional bands at: 100 kDa, 110 kDa, 145 kDa, 200 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 20 minutes
    • Flow Cytometry (Intracellular) - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
      Flow Cytometry (Intracellular) - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)

      Overlay histogram showing HeLa cells stained with ab36810 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab36810, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Western blot - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
      Western blot - Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
      All lanes : Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810) at 1/1000 dilution

      Lane 1 : Control lane
      Lane 2 : Irradiated Human fibroblasts (10 Gy gamma-irradiation)
      Lane 3 : Molecular weight marker
      Lane 4 : Peroxidated Human fibroblasts (300 µM hydrogen peroxide)
      Lane 5 : Peroxidated Human fibroblasts (1 mM hydrogen peroxide)
      Lane 6 : Peroxidated Human fibroblasts (10 mM hydrogen peroxide)

      Developed using the ECL technique.

      Predicted band size: 370 kDa
      Observed band size: 370 kDa



      Lysate loading concentration: 40µg

    文献 (75)

    发表研究结果有使用 ab36810？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab36810 被引用在 75 文献中.

    • Niwa Y  et al. Cyclin D1 Binding Protein 1 Responds to DNA Damage through the ATM-CHK2 Pathway. J Clin Med 11:N/A (2022). PubMed: 35160302
    • Cristalli C  et al. Novel Targeting of DNA Methyltransferase Activity Inhibits Ewing Sarcoma Cell Proliferation and Enhances Tumor Cell Sensitivity to DNA Damaging Drugs by Activating the DNA Damage Response. Front Endocrinol (Lausanne) 13:876602 (2022). PubMed: 35712255
    • Qin C  et al. Ataxia telangiectasia mutated: The potential negative regulator in platelet-derived growth factor-BB promoted proliferation of pulmonary arterial smooth muscle cells. Front Cardiovasc Med 9:942251 (2022). PubMed: 35990964
    • Sorteberg AL  et al. The cyclin dependent kinase inhibitor p21Cip1/Waf1 is a therapeutic target in high-risk neuroblastoma. Front Oncol 12:906194 (2022). PubMed: 36147919
    • Kriger D  et al. Alpha-actnin-4 (ACTN4) selectively affects the DNA double-strand breaks repair in non-small lung carcinoma cells. Biol Direct 17:40 (2022). PubMed: 36476259
    View all Publications for this product

    客户评价及客户问答

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    1-10 of 13 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-ATM (phospho S1981) antibody [10H11.E12]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
    Sample
    Human Cell (T98G Human brain glioblastoma)
    Specification
    T98G Human brain glioblastoma
    Permeabilization
    Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Dimitra Kalamida

    Verified customer

    提交于 Dec 12 2013

    Question

    I have worked with same catalog no abcam phospho-ATM antibody in past and I did get pATM band with the replacement stock (same catalog no but different lot no ) which * had sent me. Once I was finished with that particular vial of antibody, I ordered the new vial but that particular lot no. was not available, as we have been told, so I received phospho-ATM (ab36810 lot no ). Since then I’m having trouble detecting any bands in the western blot. I haven’t changed my protocol since last time and actually I became more critical to every step. (See attached technical sheet) I have made all of solutions fresh and did it again. Twice my western blot did not show expected pATM band.
    ) with their interpretations.
    It can’t get any worse than this: the things that used to work in the past, didn’t work at all. I don’t know what’s wrong. Any help in this matter will be greatly appreciated.

    Read More
    Answer

    Thank you for contacting Abcam.

    I am sorry about the issues you have been having with ab36818. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples. Since the current lot is not, then I would be happy to either send a replacement antibody or to process a refund.

    I can either send a new vial of ab36810 (but from a different lot) or I can send one of the following antibodies as a replacement:

    https://www.abcam.com/ATM-phospho-S1981-antibody-EP1890Y-ab81292.html
    https://www.abcam.com/ATM-phospho-S1981-antibody-ab79891.html



    Please let me know how you would like to continue and I will be happy to help resolve this issue.

    I look forward to your reply.

    Read More

    Question

    Unfortunately, even after adding secondary antibody in 5% milk instead of 5% BSA, my blot didn't show any improvement in terms of background and signals. I would like to get another vial of antibody.

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    As we have very less background in our ATM blot and that's why we thought. it could be problem with this particular vial of antibody.

    Read More
    Answer

    Thank you for your reply.


    I have another lot available. Even though this is a monoclonal antibody, it may make a difference. I would be happy to send another vial to you. Please let me know if you are agreeable to this or if you would prefer a refund.


    I look forward to your reply so that I may process your request.

    Read More

    Question

    I have done western blot again and this time i used 1:2500 dilution of primary antibody. I didn't use 1:5000 because i was skeptical that with background i would lose signals too. Also, i have done a control ATM blot from Abcam antibody which we recently purchased.
    Results and interpretations: i did see background and nonspecific bands. The band right above the 300kda looks like our pATM band. This band is seen in lanes where it should be (AdE4 mutant, Ad5 virus, Ad5 1010 mutant, and UV treatment) but it barely stands out from the non-specific bands.
    When i said the band right above 300kda is our band, it is because when i re-probed the same membrane with ATM antibody, i only observed band right above 300kda.

    I am also trying some other conditions with the remaining antibody and post you soon about that.
    Thank you so much.

    Read More
    Answer

    Thank you for your reply with this update.


    I am not sure why you are obtaining such a high degree of background with this antibody. Since your other blot is very clean, it is clearly not a technique issue such as insufficient washing.


    Can you please provide an order number for your purchase?

    Read More

    Question

    I have questions regarding the authenticity of the bands we are seeing in our blots. Please let us know whether what we are seeing is the right band?
    With this email i have attached a full questionnaire how we did our western blot as well as two blots with their interpretations/conclusions- what they mean to us.
    Please let us know your opinion.

    Read More
    Answer

    Thank you for recently completing our post complaint survey.


    I am sorry that you have been experiencing problems with this antibody. I understand you are still troubleshooting based on the protocol advice I provided. Please do not hesitate to contact me if you require further assistance. If the product continues to not work as stated on the datasheet, I am happy to offer a replacement or credit per our Abpromise guarantee.


    I wish you the best of luck with your research!

    Read More

    Question

    I am still facing difficulties in detection of pATM in western blot.

    As per your suggestion, I used 5% BSA for blocking instead of 5% milk. And I had loaded 60 ug protein in the gel.
    Results and interpretations: (Pic 1.) At the 30 sec exposure (after developing with ECL Plus) I see a lot of background. Though on a better note, I was able to see a single band above the 300Kda mark which could be pATM. But it was faint band. (Pic2). When I exposed it longer, even at 1 min, entire blot turned out black. Such background we have seen with other antibodies such as pNbs1 and pChk1 when we block with 5% BSA. So, I thought why don't I re-strip this blot and block with 5% milk and maybe I can be able to expose the film longer for a strong band.
    I restriped the same blot and used 5% milk as a blocking reagent.
    Results and interpretations: (Pic 3) At 30 sec exposure, I could lesser background and little better signals in comparison to 30 sec exposure with 5% BSA. (Pic 4) But when I exposed it longer (2min) the blot turned completely black.
    Also in both the cases, I am also observing the antibody non-specifically binding to the markers.

    I have done control blotting experiment with my secondary antibody and I haven't found any background due to secondary antibody. I could have done the control experiment with primary but I didn't to save some antibody.


    I have also done b-actin loading control experiment and found I was loading equal amount of proteins.(Pic 5)

    Now, I only have about 10 ul of antibody left. Please help me out here.

    Read More
    Answer

    Thank you for your reply.


    I am sorry to hear that you are still experiencing difficulties with this antibody in WB. I have reviewed the protocol modifications you made and the resulting blots. One thing I am not clear about is the amount of antibody you have been using. Are you still using a 1:1000 dilution or have you modified this? I have consulted with my colleagues and we all agree that it is necessary for you to decrease the amount of primary antibody being used ir order to decrease background. I would recommend testing 1:2500 and 1:5000. You should have enough antibody to test these dilutions.The last experiment will determine if the problem is with the antibody or if further optimization was needed with your samples.


    I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

    Read More

    Question

    I have questions regarding the authenticity of the bands we are seeing in our blots. Please let us know whether what we are seeing is the right band?
    With this email i have attached a full questionnaire how we did our western blot as well as two blots with their interpretations/conclusions- what they mean to us.
    Please let us know your opinion.

    Read More
    Answer

    Thank you for contacting Abcam regarding ab36810.


    I am sorry that you have been experiencing difficulties with this antibody and detecting a high level of background. I have reviewed the protocol and images you have provided and I would like to make some suggestions to improve your results with this antibody.


    You are detecting a high degree of background that must be reduced before interpreting your data. Since you are using a phospho-specific antibody, it is important that you do not use milk for blocking. I would recommend blocking with 5% BSA instead for at least 1 hour at room temperature. Finally, 100ug of total protein may be too much protein. I would recommend decreasing this as well, testing several different amounts of protein (60 and 30 ug).


    Regarding the primary antibody incubation, although 1:1000 is the recommendation on our datasheet, this will need to be optimized by the end user. I would recommend also testing 1:2000 to help reduce the background. Finally, the secondary antibody dilution appears quite low. Have you also performed a no primary control to rule out any background from the secondary antibody alone? Again the final dilution may need to be further optimized from what is present on the datasheet.


    Are you also including a loading control, such as actin? It would be useful to determine if the variations in lanes is due to uneven loading, or issues with your transfer conditions.


    I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or if these suggestions do not improve your results with this antibody.

    Read More

    Question

    Hello, The purchase order number for the pATM Ab is , and the order reference number is . A replacement antibody would be great. Thanks

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number . To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Question

    I'm afraid the 1:500 dilution did not give any signal either, even with a long exposure time. Would it be possible to get a replacement tube?

    Read More
    Answer

    Thank you for this further information.

    I am sorry that this product has not performed as stated on the datasheet. This product is guaranteed to work in WB on rat and is covered by our Abpromise for a period of six months. Since we have been unable to resolve this problem, if you have purchased this product within the last 180 days, I would be happy to either send a replacement antibody or to process a refund. Could you please send me the Abcam order confirmation number?

    Please let me know if you have any additional questions.

    Read More

    1-10 of 13 Abreviews or Q&A

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