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Anti-ATF6 antibody [1-7] (ab122897)

Key features and details

  • Mouse monoclonal [1-7] to ATF6
  • Suitable for: WB, IP, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2a

Overview

  • Product name

    Anti-ATF6 antibody [1-7]
    See all ATF6 primary antibodies

  • Description

    Mouse monoclonal [1-7] to ATF6

  • Host species

    Mouse

  • Specificity

    specific to human ATF6 alfa no cross reactivity with mouse ATF6 alfa.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment (His-tag) corresponding to Human ATF6 (N terminal). Epitope is not determined

  • Positive control

    • 293T, HeLa S3 Tet-off, 293 and HeLa cell extracts.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 6
    Constituents: 49% PBS, 50% Glycerol
  • Concentration information loading...

  • Purification notes

    ab122897 was produced from hybridoma cultured in serum-free medium and purified under mild conditions by propriety chromatography processes, then filter sterilized.

  • Clonality

    Monoclonal

  • Clone number

    1-7

  • Isotype

    IgG2a

  • Light chain type

    kappa

  • Research areas

Images

  • Western blot - Anti-ATF6 antibody [1-7] (ab122897)
    Western blot - Anti-ATF6 antibody [1-7] (ab122897)

    Lane 1: Wild type HAP1 whole cell lysate (50 µg)
    Lane 2: ATF6 knockout  HAP1 whole cell lysate (50 µg)
    Lanes 1 - 2: Merged signal (red and green). Green - ab122897 observed at 95 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab122897 was shown to specifically recact with ATF6 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when ATF6 knockout samples were used. Wild-type and ATF6 knockout samples were subjected to SDS-PAGE. Ab122897 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATF6 antibody [1-7] (ab122897)
    Immunocytochemistry/ Immunofluorescence - Anti-ATF6 antibody [1-7] (ab122897)

    Immunofluorescent analysis of HeLa cells using ab122897 at a dilution of 1/100. The cells were fixed with paraformaldehyde. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-mouse IgG at a dilution of 1/1000. The antibody staining is shown in the top panel, and the merged image with DAPI counter-staining is shown in the lower panel.

  • Western blot - Anti-ATF6 antibody [1-7] (ab122897)
    Western blot - Anti-ATF6 antibody [1-7] (ab122897)
    All lanes : Anti-ATF6 antibody [1-7] (ab122897) at 1/500 dilution

    Lane 1 : 293T cell lysate: control
    Lane 2 : 293T cell lysate: treated with 2 µg/ml tunicamycin for 7 hours
    Lane 3 : 293T cell lysate: treated with 1mM DTT for 1 hour.
    Lane 4 : HeLa S3 Tet-off cell lysate: control
    Lane 5 : HeLa S3 Tet-off cell lysate: treated with 1mM DTT for 1 hour.

    Predicted band size: 75 kDa



    Conversion of precursors pATF6(P) to pATF6(N) in DTT- or tunicamycin-treated cells. ATF6 is constitutively expressed as pATF6a(P) (~90-kDa protein), and converted to pATF6a(N) (>50-kDa protein) in ER-stressed cells.
  • Western blot - Anti-ATF6 antibody [1-7] (ab122897)
    Western blot - Anti-ATF6 antibody [1-7] (ab122897)
    All lanes : Anti-ATF6 antibody [1-7] (ab122897) at 1/500 dilution

    Lane 1 : HeLa cell extract: control
    Lane 2 : HeLa cell extract: treated with 1mM DTT for 1 hour
    Lane 3 : HeLa cell extract: treated with 2 µg/ml tunicamycin for 3 hour
    Lane 4 : HeLa cell extract: treated with 2 µg/ml tunicamycin for 7 hour
    Lane 5 : 293 cell extract: control
    Lane 6 : 293 cell extract: treated with 1mM DTT for 1 hour
    Lane 7 : 293 cell extract: treated with 2 µg/ml tunicamycin for 3 hour
    Lane 8 : 293 cell extract: treated with 2 µg/ml tunicamycin for 7 hour

    Predicted band size: 75 kDa



    Conversion of precursors pATF6(P) to pATF6(N) in DTT- or tunicamycin-treated cells. ATF6 is constitutively expressed as pATF6a(P) (~90-kDa protein), and converted to pATF6a(N) (>50-kDa protein) in ER-stressed cells.
  • Immunoprecipitation - Anti-ATF6 antibody [1-7] (ab122897)
    Immunoprecipitation - Anti-ATF6 antibody [1-7] (ab122897)

    ATF6 was detected by Western blot (Input; lanes 1, 2, 7, and 8) using ab122897 (No. 1-7). After immunoprecipitation (IP) with non-related IgG (IP; lanes 3, 4, 9, and 10) or ab122897 (No. 1-7) (IP; lanes 5, 6, 11, and 12), samples were subjected to SDS-PAGE and analyzed by Western blot using ab122897 (No. 1-7) and anti-mouse IgG antibody (light chain specific).

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