Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

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 - BSA and Azide free
  • Suitable for: ChIP, Flow Cyt, IP, ICC/IF, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ATM antibody [EPR20100] - BSA and Azide free
    See all ATM primary antibodies

  • Description

    Rabbit monoclonal [EPR20100] to ATM - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IP
    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

    Ab223533 is the carrier-free version of ab201022. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab223533 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)
    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201022).

  • Flow Cytometry - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)
    Flow Cytometry - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201022).

  • Immunoprecipitation - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)
    Immunoprecipitation - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    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 Reagent (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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201022).

  • ChIP - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)
    ChIP - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201022).

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)
    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    This ICC data was generated using the same anti-ATM antibody clone [EPR20100] in a different buffer formulation (cat# 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.

  • Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)
    Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

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