Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20100] to ATM - ChIP Grade
  • Suitable for: ICC/IF, IP, Flow Cyt, ChIP, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ATM antibody [EPR20100] - ChIP Grade
    See all ATM primary antibodies

  • Description

    Rabbit monoclonal [EPR20100] to ATM - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human testis lysate; HeLa, PC-12, RAW 264.7, 293 and SH-SY5Y whole cell lysates. ICC/IF: SH-SY5Y and HeLa cells. Flow Cyt: HEK-293 cells. IP: HEK-293 whole cell lysate. ChIP: Chromatin prepared from HCT 116 cells treated with 1mM Hydroxyurea for 16h.

  • General notes

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Western blot - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022) at 1/5000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 351 kDa
    Observed band size: 351 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • Western blot - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Western blot - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    All lanes : Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022) at 1/1000 dilution

    Lane 1 : Human testis lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 3 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 4 : 293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 5 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 351 kDa
    Observed band size: 351 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 3 minutes; Lane 3: 30 seconds; Lane 4: 5 seconds; Lane 5: 1 second

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATM with ab201022 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling ATM with ab201022 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on SH-SY5Y cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow Cytometry - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Flow Cytometry - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling ATM with ab201022 at 1/800 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Immunoprecipitation - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

    ATM was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate with ab201022 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab201022 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293 whole cell lysate, 10 μg (Input).

    Lane 2: ab201022 IP in HEK-293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201022 in HEK-293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    ATM cleavage has been documented previously and the fragment pattern is consistent with what has been described in the literature PMID:16849690

  • ChIP - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    ChIP - Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

    Chromatin was prepared from HCT 116 (Human colorectal carcinoma cell line) cells treated with 1mM Hydroxyurea for 16h and non-treated according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab201022 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow).  The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)
    Anti-ATM antibody [EPR20100] - ChIP Grade (ab201022)

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