Overview

  • Product nameAnti-ATF6 antibody
    See all ATF6 primary antibodies
  • Description
    Rabbit polyclonal to ATF6
  • Tested applicationsSuitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Human ATF6.

    (Peptide available as ab92653.)

  • Positive control
    • This antibody gave a positive signal in MDA-MB-361 Whole Cell Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab83504 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 75,90,100 kDa (predicted molecular weight: 75 kDa).

Target

  • FunctionTranscription factor that acts during endoplasmic reticulum stress by activating unfolded protein response target genes. Binds DNA on the 5'-CCAC[GA]-3'half of the ER stress response element (ERSE) (5'-CCAAT-N(9)-CCAC[GA]-3') and of ERSE II (5'-ATTGG-N-CCACG-3'). Binding to ERSE requires binding of NF-Y to ERSE. Could also be involved in activation of transcription by the serum response factor.
  • Tissue specificityUbiquitous.
  • Sequence similaritiesBelongs to the bZIP family. ATF subfamily.
    Contains 1 bZIP domain.
  • DomainThe basic domain functions as a nuclear localization signal.
    The basic leucine-zipper domain is sufficient for association with the NF-Y trimer and binding to ERSE.
  • Post-translational
    modifications
    During unfolded protein response an approximative 50 kDa fragment containing the cytoplasmic transcription factor domain is released by proteolysis. The cleavage seems to be performed sequentially by site-1 and site-2 proteases.
    N-glycosylated. The glycosylation status may serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the unfolded protein response (UPR).
    Phosphorylated in vitro by MAPK14/P38MAPK.
  • Cellular localizationEndoplasmic reticulum membrane and Nucleus. Under ER stress the cleaved N-terminal cytoplasmic domain translocates into the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Activating transcription factor 6 alpha antibody
    • Activating transcription factor 6 antibody
    • ATF 6 antibody
    • ATF6 alpha antibody
    • ATF6 antibody
    • ATF6-alpha antibody
    • ATF6A antibody
    • ATF6A_HUMAN antibody
    • cAMP dependent transcription factor ATF 6 alpha antibody
    • cAMP-dependent transcription factor ATF-6 alpha antibody
    • Cyclic AMP dependent transcription factor ATF 6 alpha antibody
    • DKFZp686P2194 antibody
    • ESTM49 antibody
    • FLJ21663 antibody
    • Processed cyclic AMP dependent transcription factor ATF 6 alpha antibody
    • Processed cyclic AMP-dependent transcription factor ATF-6 alpha antibody
    see all

Anti-ATF6 antibody images

  • All lanes : Anti-ATF6 antibody (ab83504) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate (untreated)
    Lane 2 : HeLa Whole Cell Lysate (treated with 2.37 uM Tunicamycin)
    Lane 3 : HeLa Whole Cell Lysate (treated with 23.7 uM Tunicamycin)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 75 kDa
    Observed band size : 75,90 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa (possible glycosylated form).

    Exposure time : 8 minutes

    ATF6 has a predicted molecular weight of 75 kDa; however it has a number of potential glycosylation sites which may affect the migration of the protein (SwissProt data). We believe the bands observed at 75 and 100 kDa correspond to the full length nonglycosylated and glycosylated forms of ATF6.  Following Tunicamycin treatment to inhibit N-linked glycosylation we see the appearance of a partially deglycosylated form at 90 kDa.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab83504 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • Anti-ATF6 antibody (ab83504) at 1 µg/ml + MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 75 kDa
    Observed band size : 75,78 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60 kDa,90 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab83504 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab83504, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-ATF6 antibody (ab83504)

ab83504 has not yet been referenced specifically in any publications.

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