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Western blot - ATF6 antibody (ab37149)
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Immunocytochemistry/ Immunofluorescence - ATF6 antibody (ab37149)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-ATF6 antibody(ab37149)
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Product Name
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ATF6 antibody
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See all ATF6 antibodies (6)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to ATF6
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Immunogen
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Synthetic peptide corresponding to 17 amino acids near to the N terminus of the human ATF6 protein.
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Reacts with
(species key)
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Hu, Ms
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Specificity
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Based on 100% sequence alignment, this antibody detects ATF6 isoform A (alpha). The antibody is predicted to recognize the 50kDa fragment as the it recognizes a peptide sequence from near the amino terminus. We have not tested stressed cells to see if the 50kDa cleavage product can be detected.
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Tested applications
(see key)
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ICC/IF, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC/IF: Use at a concentration of 10 µg/ml.
IHC-P: Use at a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 75 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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MDA MB 361 cell lysate.
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Cellular localization
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Endoplasmic reticulum: Type II membrane protein in the endoplasmic reticulum. Under ER stress the cleaved N terminal cytoplasmic domain translocates into the nucleus.
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Research areas
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Chromatin and Nuclear Signaling >> Transcription >> Domain Families >> HLH / Leucine Zipper >> Leucine Zipper
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Relevance
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ATF6 is a transcription factor that acts during endoplasmic reticulum stress by activating unfolded protein response target genes. It 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. ATF6 could also be involved in activation of transcription by the serum response factor. ATF6 exists as a homodimer and heterodimer with ATF6 beta. The dimer interacts with the nuclear transcription factor Y (NF-Y) trimer through direct binding to NF-Y subunit C (NF-YC). It Interacts also with the transcription factors GTF2I, YY1 and SRF. Under ER stress the cleaved N-terminal cytoplasmic domain translocates into the nucleus. The basic domain of ATF6 functions as a nuclear localization signal and the basic leucine zipper domain is sufficient for association with the NF-Y trimer and binding to ERSE. During the unfolded protein response an approximately 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. ATF6 is N glycosylated, phosphorylated in vitro by MAPK14/P38MAPK and belongs to the bZIP family.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: PBS Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Purification notes
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Immunoaffinity purified
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Store at +4°C.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this ATF6 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for ATF6 antibody images, references, products related to ab37149 and other tools.
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ATF6 antibody images:
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 Western blot - ATF6 antibody (ab37149)
Lane 1 : ATF6 antibody (ab37149) at 0.5 µg/ml
Lane 2 : ATF6 antibody (ab37149) at 1 µg/ml
Lane 1 : MDA MB 361 cell lysate
Lane 2 : MDA MB 361 cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
goat anti-rabbit HRP at 1/10000 dilution
Predicted band size : 75 kDa
Observed band size : 85 kDa (why is the actual band size different from the predicted?)
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Immunocytochemistry/ Immunofluorescence - ATF6 antibody (ab37149)
ab37149 staining human Hella cells by ICC/IF. Cells were PFA fixed and permeabilized for 10 minutes in 0.1% Triton X-100 prior to blocking in 3% BSA for 15 minutes at 25°C. The primary antibody was diluted 1/200 and incubated with the sample for 1 hour. The secondary antibody used was Alexa Fluor®568 goat anti-rabbit diluted at 1/1000. This image is courtesy of an Abreview submitted by Dr Dan Tattersall
See Abreview
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-ATF6 antibody(ab37149)
Ab37149 staining human normal pancreas. Staining is localised to the cytoplasm.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the DAKO 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
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References for ATF6 antibody (ab37149)
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This product has been used in:
Jian B et al. Activation of endoplasmic reticulum stress response following trauma-hemorrhage. Biochim Biophys Acta 1782:621-6 (2008). PubMed: 18801427
If you publish research using ab37149 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to ATF6
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ATF6 antibody - more information
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ATF6 antibody - other tools:
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Contact Abcam with a Technical Enquiry about ATF6 antibody |
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