Recombinant Anti-ATF6 antibody [EPR22690-84] - ChIP Grade (ab227830)
![Western blot - Anti-ATF6 antibody [EPR22690-84] (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-355787-227830WB1anti-atf6-antibody-epr22690-84-western-blot-hap1-human.jpg "Western blot - Anti-ATF6 antibody [EPR22690-84] (ab227830)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22690-84] to ATF6 - ChIP Grade
- Suitable for: WB, IHC-P, ChIP, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-ATF6 antibody [EPR22690-84] - ChIP Grade
See all ATF6 primary antibodies -
Description
Rabbit monoclonal [EPR22690-84] to ATF6 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ChIP, IPmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1 and HeLa whole cell lysates. IHC-P: Human kidney tissue. IP: HeLa whole cell lysate. ChIP: Chromatin prepared from RAW 264.7 (treated with tunicamycin) and HeLa (treated with thapsigargin) cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22690-84 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227830 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) |
1/1000. Predicted molecular weight: 74 kDa.
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| IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| ChIP |
Use 5 µg for 25 µg of chromatin.
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| IP |
1/30.
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| Notes |
|---|
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WB
1/1000. Predicted molecular weight: 74 kDa. |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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ChIP
Use 5 µg for 25 µg of chromatin. |
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IP
1/30. |
Target
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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
modificationsDuring 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
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Database links
- Entrez Gene: 22926 Human
- Entrez Gene: 226641 Mouse
- Omim: 605537 Human
- SwissProt: P18850 Human
- Unigene: 492740 Human
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Alternative names
- Activating transcription factor 6 alpha antibody
- Activating transcription factor 6 antibody
- ATF 6 antibody
see all
Images
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Western blot - Anti-ATF6 antibody [EPR22690-84] (ab227830)All lanes : Anti-ATF6 antibody [EPR22690-84] - ChIP Grade (ab227830) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : ATF6 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 74 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?ab227830 was shown to specifically react with ATF6 in wild-type HAP1 cells as signal was lost in ATF6 knockout cells. Wild-type and ATF6 knockout samples were subjected to SDS-PAGE. ab227830 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging.Exposure time 62 seconds.Blocking/Diluting buffer and concentration: 5% NFDM/TBST. -
![Western blot - Anti-ATF6 antibody [EPR22690-84] (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-355788-227830WB2anti-atf6-antibody-epr22690-84-western-blot-hela-human.jpg)
Western blot - Anti-ATF6 antibody [EPR22690-84] (ab227830)All lanes : Anti-ATF6 antibody [EPR22690-84] - ChIP Grade (ab227830) at 1/1000 dilution
Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa treated with 1 µ? thapsigargin for 1 hour, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 74 kDa
Observed band size: 50,90 kDa why is the actual band size different from the predicted?ATF6 is cleaved upon ER stress and the molecular weight observed is consistent with what has been described in the literature (PMID: 25149687; 11163209).Exposure time 3 minutes.Blocking and diluting buffer and concentration: 5% NFDM/TBST. -
![Immunoprecipitation - Anti-ATF6 antibody [EPR22690-84] (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-355786-227830IP1anti-atf6-antibody-epr22690-84-immunoprecipitation-hela-human.jpg)
Immunoprecipitation - Anti-ATF6 antibody [EPR22690-84] (ab227830)ATF6 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab227830 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab227830 1/1000 dilution (0.5 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2: ab254324 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227830 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 min.
Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody [EPR22690-84] (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-355785-227830IHC1anti-atf6-antibody-epr22690-84-immunohistochemistry-kidney-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody [EPR22690-84] (ab227830)Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling ATF6 with ab227830 at 1/1000 dilution (0.566 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney (PMID: 25725420) is observed. The section was incubated with ab227830 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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![ChIP - Anti-ATF6 antibody [EPR22690-84] (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-2-anti-atf6-antibody-epr22690-84-chromatin-immunoprecipitation-hela-human.jpg)
ChIP - Anti-ATF6 antibody [EPR22690-84] (ab227830)Chromatin was prepared from HeLa cells treated with thapsigargin (1 uM, 1 h) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30 mins and then formaldehyde for 10 min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab227830 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are from paper PMID: 17535801.
*https://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
![ChIP - Anti-ATF6 antibody [EPR22690-84] (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-1-anti-atf6-antibody-epr22690-84-chromatin-immunoprecipitation-raw-2647-mouse.jpg)
ChIP - Anti-ATF6 antibody [EPR22690-84] (ab227830)Chromatin was prepared from RAW 264.7 cells treated with tunicamycin (5 ug/ml, 4 h) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30 mins and then formaldehyde for 10 min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab227830 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are from paper PMCID: PMC5179193.
*https://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
![Anti-ATF6 antibody [EPR22690-84] - ChIP Grade (ab227830)](https://www.abcam.com/ps/products/227/ab227830/Images/ab227830-7-benefits-of-recombinant-antibodies.png)
Anti-ATF6 antibody [EPR22690-84] - ChIP Grade (ab227830)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (9)
ab227830 has been referenced in 9 publications.
- Zhang H et al. Rutaecarpine ameliorates lipopolysaccharide-induced BEAS-2B cell injury through inhibition of endoplasmic reticulum stress via activation of the AMPK/SIRT1 signaling pathway. Exp Ther Med 23:373 (2022). PubMed: 35495603
- Fan H et al. α-Linolenic Acid Suppresses Proliferation and Invasion in Osteosarcoma Cells via Inhibiting Fatty Acid Synthase. Molecules 27:N/A (2022). PubMed: 35566090
- Wang L et al. Curcumin Increased the Sensitivity of Non-Small-Cell Lung Cancer to Cisplatin through the Endoplasmic Reticulum Stress Pathway. Evid Based Complement Alternat Med 2022:6886366 (2022). PubMed: 35754693
- Xia H et al. Selenoprotein K Is Essential for the Migration and Phagocytosis of Immature Dendritic Cells. Antioxidants (Basel) 11:N/A (2022). PubMed: 35883755
- Wufuer R et al. Distinct Roles of Nrf1 and Nrf2 in Monitoring the Reductive Stress Response to Dithiothreitol (DTT). Antioxidants (Basel) 11:N/A (2022). PubMed: 36009254
- Chen D et al. High-fat diet aggravates colitis via mesenteric adipose tissue derived exosome metastasis-associated lung adenocarcinoma transcript 1. World J Gastroenterol 28:3838-3853 (2022). PubMed: 36157545
- Ma M et al. ATF6 aggravates angiogenesis-osteogenesis coupling during ankylosing spondylitis by mediating FGF2 expression in chondrocytes. iScience 24:102791 (2021). PubMed: 34296071
- Ko P et al. Microtubule Acetylation Controls MDA-MB-231 Breast Cancer Cell Invasion through the Modulation of Endoplasmic Reticulum Stress. Int J Mol Sci 22:N/A (2021). PubMed: 34199510
- Song Z et al. Low DAPK1 expression correlates with poor prognosis and sunitinib resistance in clear cell renal cell carcinoma. Aging (Albany NY) 13:1842-1858 (2020). PubMed: 33201837