ATF6 Antibody | Affinity Biosciences

Product:	ATF6 Antibody
Catalog:	DF6009
Description:	Rabbit polyclonal antibody to ATF6
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Zebrafish, Horse, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.:	50~75kD(cleaved),90~100kD(full); 75kD(Calculated).
Uniprot:	P18850
RRID:	AB_2833019

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Zebrafish(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(100%)

Clonality:

Polyclonal

Specificity:

ATF6 Antibody detects endogenous levels of total ATF6.

RRID:

AB_2833019
Cite Format: Affinity Biosciences Cat# DF6009, RRID:AB_2833019.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

Activating transcription factor 6 alpha; Activating transcription factor 6; ATF 6; ATF6 alpha; ATF6; ATF6-alpha; ATF6A; ATF6A_HUMAN; cAMP dependent transcription factor ATF 6 alpha; cAMP-dependent transcription factor ATF-6 alpha; Cyclic AMP dependent transcription factor ATF 6 alpha; DKFZp686P2194; ESTM49; FLJ21663; Processed cyclic AMP dependent transcription factor ATF 6 alpha; Processed cyclic AMP-dependent transcription factor ATF-6 alpha;

Immunogens

Immunogen:

Uniprot:

P18850(ATF6A_HUMAN) >>Visit HPA database.

Gene(ID):

ATF6(22926)

Expression:

P18850 ATF6A_HUMAN:

Ubiquitous.

Description:

This gene encodes a transcription factor that activates target genes for the unfolded protein response (UPR) during endoplasmic reticulum (ER) stress. Although it is a transcription factor, this protein is unusual in that it is synthesized as a transmembrane protein that is embedded in the ER. It functions as an ER stress sensor/transducer, and following ER stress-induced proteolysis, it functions as a nuclear transcription factor via a cis-acting ER stress response element (ERSE) that is present in the promoters of genes encoding ER chaperones. This protein has been identified as a survival factor for quiescent but not proliferative squamous carcinoma cells. There have been conflicting reports about the association of polymorphisms in this gene with diabetes in different populations, but another polymorphism has been associated with increased plasma cholesterol levels. This gene is also thought to be a potential therapeutic target for cystic fibrosis. [provided by RefSeq, Aug 2011]

Sequence:

P18850 ATF6A_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MGEPAGVAGTMESPFSPGLFHRLDEDWDSALFAELGYFTDTDELQLEAANETYENNFDNLDFDLDLMPWESDIWDINNQICTVKDIKAEPQPLSPASSSYSVSSPRSVDSYSSTQHVPEELDLSSSSQMSPLSLYGENSNSLSSAEPLKEDKPVTGPRNKTENGLTPKKKIQVNSKPSIQPKPLLLPAAPKTQTNSSVPAKTIIIQTVPTLMPLAKQQPIISLQPAPTKGQTVLLSQPTVVQLQAPGVLPSAQPVLAVAGGVTQLPNHVVNVVPAPSANSPVNGKLSVTKPVLQSTMRNVGSDIAVLRRQQRMIKNRESACQSRKKKKEYMLGLEARLKAALSENEQLKKENGTLKRQLDEVVSENQRLKVPSPKRRVVCVMIVLAFIILNYGPMSMLEQDSRRMNPSVSPANQRRHLLGFSAKEAQDTSDGIIQKNSYRYDHSVSNDKALMVLTEEPLLYIPPPPCQPLINTTESLRLNHELRGWVHRHEVERTKSRRMTNNQQKTRILQGALEQGSNSQLMAVQYTETTSSISRNSGSELQVYYASPRSYQDFFEAIRRRGDTFYVVSFRRDHLLLPATTHNKTTRPKMSIVLPAININENVINGQDYEVMMQIDCQVMDTRILHIKSSSVPPYLRDQQRNQTNTFFGSPPAATEATHVVSTIPESLQ

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Horse

100

Sheep

100

Dog

100

Xenopus

100

Zebrafish

100

Rabbit

100

Bovine

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P18850 As Substrate

P18850

Site	PTM Type	Enzyme	Source
S16	Phosphorylation		Uniprot
K87	Sumoylation		Uniprot
K87	Ubiquitination		Uniprot
S94	Phosphorylation		Uniprot
Y100	Phosphorylation		Uniprot
S104	Phosphorylation		Uniprot
K152	Sumoylation		Uniprot
T166	Phosphorylation	Q16539 (MAPK14)	Uniprot
K176	Acetylation		Uniprot
K176	Sumoylation		Uniprot
K176	Ubiquitination		Uniprot
K182	Sumoylation		Uniprot
K191	Sumoylation		Uniprot
K201	Ubiquitination		Uniprot
K216	Ubiquitination		Uniprot
K290	Sumoylation		Uniprot
K290	Ubiquitination		Uniprot
T296	Phosphorylation		Uniprot
Y330	Phosphorylation		Uniprot
K339	Ubiquitination		Uniprot
K349	Ubiquitination		Uniprot
S373	Phosphorylation		Uniprot
S410	Phosphorylation		Uniprot
K424	Ubiquitination		Uniprot
K436	Ubiquitination		Uniprot
N472	N-Glycosylation		Uniprot
K506	Ubiquitination		Uniprot
T565	Phosphorylation		Uniprot
N584	N-Glycosylation		Uniprot
K629	Ubiquitination		Uniprot
S632	Phosphorylation		Uniprot
N643	N-Glycosylation		Uniprot

Research Backgrounds

P18850_ATF6A_HUMAN

Function:

Transmembrane glycoprotein of the endoplasmic reticulum that functions as a transcription activator and initiates the unfolded protein response (UPR) during endoplasmic reticulum stress. Cleaved upon ER stress, the N-terminal processed cyclic AMP-dependent transcription factor ATF-6 alpha translocates to the nucleus where it activates transcription of genes involved in the UPR. 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. May play a role in foveal development and cone function in the retina.

PTMs:

During unfolded protein response, a fragment of approximately 50 kDa 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.

Subcellular Location:

Endoplasmic reticulum membrane>Single-pass type II membrane protein.

Nucleus.
Note: Under ER stress the cleaved N-terminal cytoplasmic domain translocates into the nucleus. THBS4 promotes its nuclear shuttling.

Tissue Specificity:

Ubiquitous.

Subunit Structure:

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). Interacts also with the transcription factors GTF2I, YY1 and SRF. Interacts (via lumenal domain) with THBS1 (By similarity). Interacts with THBS4 (via EGF-like 3; calcium-binding domain) which facilitates its processing, activation and nuclear translocation. Interacts with XBP1 isoform 2; the interaction occurs in a ER stress-dependent manner.

Family&Domains:

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.

Belongs to the bZIP family. ATF subfamily.

Research Fields

· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum. (View pathway)

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

References

1). Branched chain amino acids exacerbate myocardial ischemia/reperfusion vulnerability via enhancing GCN2/ATF6/PPAR-α pathway-dependent fatty acid oxidation. Theranostics (PubMed: 32373236) [IF=12.4]

Application: WB Species: mouse Sample: Heart

Figure 6. BCAA increase PPAR-α expression in a GCN2/ATF6 pathway-dependent manner. (A) Expression of p-GCN2, GCN2 and ATF6 in the presence of increasing concentrations of BCAA (0, 0.429 mM, 0.858 mM, 1.716 mM, 3.432 mM) by western blotting (n=6). (B) Expression of p-GCN2, GCN2 and ATF6 in the presence of increasing concentrations of BCKA (0, 0.429 mM, 0.858 mM, 1.716 mM, 3.432 mM) by western blotting (n=6). BCKA mixture is composed of αKIC, αKIV and αKMV (weight ratio, αKIC: αKIV: αKMV= 2:1:1). (C) NRVMs were treated with control siRNA and ATF6 siRNA. 48 h after transfection, expression of ATF6 was determined by western blotting (n=4). (D-E) ATF6 siRNA transferred NRVMs were treated with or without BCAA (3.432 mM) (n=6). (D) PPAR-α expression was determined by western blotting. (E) Expression of Acaa2, Acadm, Cd36 and Cpt1b by real-time PCR. (F-G) ATF6 siRNA transferred NRVMs were treated with or without BCKA (3.432 mM) (n=6). (F) PPAR-α expression was determined by western blotting. (G) Expression of Acaa2, Acadm, Cd36 and Cpt1b by real-time PCR. (C) Data were analyzed by Student’s t test. (A-B) and (D-G) Data were analyzed by one-way ANOVA, followed by a Bonferroni post-hoc test. * P<0.05, ** P<0.01. All values are presented as mean ± SEM.

2). Zonisamide, an antiepileptic drug, alleviates diabetic cardiomyopathy by inhibiting endoplasmic reticulum stress. Acta Pharmacologica Sinica (PubMed: 32647341) [IF=8.2]

Application: WB Species: mice Sample: NRCMs

Fig. 6 Zonisamide alleviates HG-induced cardiac hypertrophy and apoptosis in cultured primary neonatal rat cardiomyocytes (NRCMs) via suppression of activated ER stress. NRCMs were pretreated with 5 mM 4-PBA (an inhibitor of ERS) or 10 ng/mL tunicamycin (Tm, an ERS inducer) for 2 h and then exposed to glucose (33 mM) in the presence or absence of ZNS (3 μM) for 24 h. a–b Representative and quantitative images showing the protein expression of ERS markers, including GRP78, XBP-1s, ATF6, p-PERK, PERK, ATF4, CHOP, and Hrd1. c Immunofluorescence staining of cardiomyocytes with phalloidin (red) and cell nuclei with DAPI (blue), Scale bar = 50 μm. d Quantitative analysis of cell surface area by ImageJ software. e–f Representative Western blotting and analysis of Bax and Bcl-2 expression. g–h Representative and quantitative images of GRP78, ATF6, p-PERK, PERK, ATF4, and CHOP expression. All values are the fold changes normalized to their control group. The results are presented as the means ± SEM (n = 6). *P < 0.05, **P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. HG; $P < 0.05, $$P < 0.01 vs. HG + ZNS.

3). Oleic acid protects saturated fatty acid mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis. LIFE SCIENCES (PubMed: 29709653) [IF=6.1]

4). The potential immunotoxicity of fine particulate matter based on SD rat spleen. ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH (PubMed: 31218585) [IF=5.8]

Application: WB Species: rat Sample: spleen

Fig. 3 | PM2.5 induced ERS in spleen of SD rats. a, b The levels of spleen GRP78 and ATF6 mRNA were measured using qRT-PCR by summer and winter PM2.5 treatment in SD rat. c, dThe levels of spleen GRP78 and ATF6 protein were measured using western blotting in summer PM2.5 treatment in SD rats and quantification of analysis

5). Chrysin ameliorates synovitis and fibrosis of osteoarthritic fibroblast-like synoviocytes in rats through PERK/TXNIP/NLRP3 signaling. Frontiers in Pharmacology (PubMed: 37021049) [IF=5.6]

6). Peroxisome Proliferator-Activated Receptor-Gamma Reduces ER Stress and Inflammation via Targeting NGBR Expression. Frontiers in Pharmacology (PubMed: 35111067) [IF=5.6]

Application: WB Species: Human Sample: HUVEC cells

FIGURE 6 PPARγ reduces tunicamycin-induced ER stress by regulating NGBR. (A–C) HUVEC cells in a six-well plate were transfected with control siRNA (NC siRNA) or NGBR siRNA for 24 h in serum-free medium, followed by switching the cells into complete medium to culture for another 24 h. After treatment with rosiglitazone (10 μM) for 12 h, the cells were treated with tunicamycin (0.5 μg/ml) with or without rosiglitazone for another 12 h. Expression of CHOP, BIP, NGBR p-PERK, p-IRE1α and c-ATF6 protein was determined by Western blot (A, B). Expression of CHOP, BIP and NGBR mRNA was determined by qPCR (C). Values were expressed as means ± SD. *p < 0.05; **p < 0.01; ns, not significant (n = 3). (D) liver total proteins were collected from Figure 4. Expression of BIP and CHOP in mouse liver was determined by Western blot. Values were expressed as means ± SD. *p < 0.05; **p < 0.01; ns, not significant (n = 3).

7). Protective Effect of Patchouli Alcohol Against High-Fat Diet Induced Hepatic Steatosis by Alleviating Endoplasmic Reticulum Stress and Regulating VLDL Metabolism in Rats. Frontiers in Pharmacology (PubMed: 31632274) [IF=5.6]

Application: WB Species: rat Sample: liver

FIGURE 4 | PA treatment attenuated HFD-induced ER stress in rats. (A) Representative immunoreactive bands of GRP78, PERK, p-PERK, IRE1α, p-IRE1α, and ATF6

8). Fine particulate matter promotes airway inflammation and mucin production by activating endoplasmic reticulum stress and the IRE1α/NOD1/NF‑κB pathway. International Journal of Molecular Medicine (PubMed: 37654182) [IF=5.4]

Application: WB Species: Human Sample: HBE135-E6E7 cells

Figure 2 Expression of endoplasmic reticulum stress-related proteins in HBE135-E6E7 cells. Cells were exposed to 100 μg/ml PM2.5, or treated with 5 mmol/l 4-PBA prior to PM2.5 exposure. The levels of the aforementioned proteins were measured using western blot analysis. (A) The relative protein expression levels of GRP78 and CHOP are depicted as the ratio of each to β-actin. The relative p-IRE1α protein expression levels are presented as the ratio of p-IRE1α to IRE1α; β-actin blots were used as the loading control. (B) The same method was used to indicate the relative expression levels of NOD1 and ATF6 proteins. (C) Relative p-PERK protein expression levels. Data are presented as the mean ± SD (n=3 repeats per group). *P

9). Mesenchymal stromal cells protect hepatocytes from lipotoxicity through alleviation of endoplasmic reticulum stress by restoring SERCA activity. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (PubMed: 33591626) [IF=5.3]

Application: WB Species: human Sample: HepG2 cells

FIGURE 5 |MSCs alleviated PA-induced ER stress in HepG2 cells. The morphology of the ER in the HepG2 cells was observed by electron microscopy (A). The mRNA expression and proteins levels of ER stress makers and the protein levels of BiP, ATF6/4, p-eIF2α and CHOP in HepG2 cells were measured after 24 h (B, C).

Application: WB Species: Human Sample: HepG2 cells

FIGURE 5 MSCs alleviated PA‐induced ER stress in HepG2 cells. The morphology of the ER in the HepG2 cells was observed by electron microscopy (A). The mRNA expression and proteins levels of ER stress makers and the protein levels of BiP, ATF6/4, p‐eIF2α and CHOP in HepG2 cells were measured after 24 h (B, C). Representative Fluo‐4AM ratio images of cytosolic calcium in HepG2 cells after 24 h, and quantification of the relative fluorescence intensity (D). The intracellular calcium release was detected by flow cytometry (E). The protein and mRNA expression levels of SERCA in HepG2 cells after PA exposure for 24 h with or without MSC coculture (F, G). Measurement of the SERCA activity in HepG2 cells at 24 h after exposure (H). The protein and mRNA expression levels of SERCA in the livers of rats (I, J). Results are presented as means ± SD from three independent experiments, *P < .05 vs the control or BSA group; # P < .05 vs the PA or HFD group

10). Benzo[a]pyrene treatment modulates Nrf2/Keap1 axis and changes the metabolic profile in rat lung cancer. Chemico-Biological Interactions (PubMed: 36736873) [IF=5.1]

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ATF6 Antibody - #DF6009