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

    Suitable for: WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse
  • 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

    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

Applications

Our Abpromise guarantee covers the use of ab122897 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
WB 1/500 - 1/1000. Predicted molecular weight: 75 kDa.

If clear result not obtained Immunoprecipitation may help

IP Use at an assay dependent concentration.
ICC/IF 1/100.

Target

  • Function

    Transcription 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 specificity

    Ubiquitous.
  • Sequence similarities

    Belongs to the bZIP family. ATF subfamily.
    Contains 1 bZIP domain.
  • Domain

    The 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 localization

    Endoplasmic 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

Images

  • 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.

  • 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.

  • 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.
  • 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.
  • 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).

References

This product has been referenced in:

  • Westergard T  et al. Repeat-associated non-AUG translation in C9orf72-ALS/FTD is driven by neuronal excitation and stress. EMBO Mol Med 11:N/A (2019). Read more (PubMed: 30617154) »
  • Nguyen P  et al. GIV/Girdin promotes cell survival during endoplasmic reticulum stress. Mol Cell Biochem 453:79-88 (2019). Read more (PubMed: 30145643) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

This product is known to not work in this application or species.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mice Hepatocytes)
Gel Running Conditions
Reduced Denaturing (4-15% SDS-PAGE)
Loading amount
35 µg
Specification
Mice Hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Armen Petrosyan

Verified customer

Submitted Feb 20 2019

Application
Flow Cytometry
Sample
Human Cell (Human primary monocyte-derived macrophages)
Permeabilization
Yes - 0.5% Triton X100
Gating Strategy
Gated on the non-debris population
Specification
Human primary monocyte-derived macrophages
Preparation
Cell harvesting/tissue preparation method: Cells lifted by Tryple express
Sample buffer: FACS buffer (PBS with BSA and EDTA) used as the sample buffer
Fixation
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 16 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293)
Permeabilization
Yes - 0.3% Triton X-100 in blocking buffer
Specification
HEK293
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 05 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
human hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 28 2017

Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
35 µg
Specification
Rat hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Armen Petrosyan

Verified customer

Submitted Oct 20 2017

Application
Western blot
Sample
Human Tissue lysate - whole (WIF-B cells, the hepatoma-derived hybrid cell line)
Gel Running Conditions
Reduced Denaturing (6% SDS-PAGE)
Loading amount
30 µg
Specification
WIF-B cells, the hepatoma-derived hybrid cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Armen Petrosyan

Verified customer

Submitted Dec 08 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Nov 30 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (DU145)
Permeabilization
Yes - 0.2% Triton X-100
Specification
DU145
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Oct 31 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (HBEC)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
50 µg
Treatment
glucose deprivation
Specification
HBEC
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 08 2016

Application
Western blot
Loading amount
80 µg
Gel Running Conditions
Reduced Denaturing
Sample
African Green Monkey Cell lysate - whole cell (Cox7)
Specification
Cox7
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1.25% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 24 2014

1-10 of 11 Abreviews or Q&A

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