ATF4 Antibody - #DF6008

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Product: ATF4 Antibody
Catalog: DF6008
Description: Rabbit polyclonal antibody to ATF4
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Dog, Chicken
Mol.Wt.: 39~49kD; 39kD(Calculated).
Uniprot: P18848
RRID: AB_2833025

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 100ul $280 In stock
 200ul $350 In stock

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Related Downloads
MS Report
HPLC Report
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%), Bovine(100%), Horse(80%), Sheep(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
ATF4 Antibody detects endogenous levels of total ATF4.
RRID:
AB_2833025
Cite Format: Affinity Biosciences Cat# DF6008, RRID:AB_2833025.
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 4; ATF 4; ATF4; ATF4 protein; ATF4_HUMAN; cAMP-dependent transcription factor ATF-4; cAMP-responsive element-binding protein 2; CREB 2; CREB-2; CREB2; Cyclic AMP dependent transcription factor ATF 4; Cyclic AMP response element binding protein 2; Cyclic AMP-dependent transcription factor ATF-4; Cyclic AMP-responsive element-binding protein 2; DNA binding protein TAXREB67; DNA-binding protein TAXREB67; Tax Responsive Enhancer Element B67; Tax-responsive enhancer element-binding protein 67; TaxREB67; TXREB;

Immunogens

Immunogen:
Uniprot:

P18848(ATF4_HUMAN) >>Visit HPA database.

Gene(ID):
ATF4(468)
Description:
ATF4 is a transcription factor, that accumulates predominantly in osteoblasts, where it regulates terminal osteoblast differentiation and bone formation. As a basic leucine-zipper (bZip) transcription factor, ATF4 can regulate amino acid metabolism, cellular redox state, and anti-stress responses. It also regulates age-related and diet-induced obesity and glucose homeostasis in mammals, and has conserved metabolic functions in flies. Due to its location at chromosome 22q13, a region linked to schizophrenia, ATF4 is considered as a positional candidate gene for schizophrenia. Otherwise, since ATF4 is induced by tumourmicroenvironmental factors, and regulates processes relevant to cancer progression, it might serve as a potential therapeutic target in cancer. This antibody is a rabbit polyclonal antibody raised against full length human ATF4 antigen.
Sequence:
MTEMSFLSSEVLVGDLMSPFDQSGLGAEESLGLLDDYLEVAKHFKPHGFSSDKAKAGSSEWLAVDGLVSPSNNSKEDAFSGTDWMLEKMDLKEFDLDALLGIDDLETMPDDLLTTLDDTCDLFAPLVQETNKQPPQTVNPIGHLPESLTKPDQVAPFTFLQPLPLSPGVLSSTPDHSFSLELGSEVDITEGDRKPDYTAYVAMIPQCIKEEDTPSDNDSGICMSPESYLGSPQHSPSTRGSPNRSLPSPGVLCGSARPKPYDPPGEKMVAAKVKGEKLDKKLKKMEQNKTAATRYRQKKRAEQEALTGECKELEKKNEALKERADSLAKEIQYLKDLIEEVRKARGKKRVP

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
Bovine
100
Sheep
100
Dog
100
Chicken
100
Horse
80
Xenopus
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P18848 As Substrate

Site PTM Type Enzyme
K45 Sumoylation
K45 Ubiquitination
K53 Sumoylation
K53 Ubiquitination
K55 Ubiquitination
S69 Phosphorylation
K75 Ubiquitination
K88 Ubiquitination
T107 Phosphorylation P07949 (RET)
T114 Phosphorylation P07949 (RET)
T115 Phosphorylation P07949 (RET)
T119 Phosphorylation P07949 (RET)
S215 Phosphorylation
S219 Phosphorylation
S224 Phosphorylation
S245 Phosphorylation P51812 (RPS6KA3)
S248 Phosphorylation
K259 Ubiquitination
K267 Sumoylation
K267 Ubiquitination
K277 Ubiquitination
T293 Phosphorylation
Y295 Phosphorylation
K299 Ubiquitination
K311 Acetylation PR:000007102 (EP300)
K329 Ubiquitination
K335 Ubiquitination
K343 Acetylation
K348 Acetylation

Research Backgrounds

Function:

Transcriptional activator. Binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Cooperates with FOXO1 in osteoblasts to regulate glucose homeostasis through suppression of beta-cell production and decrease in insulin production (By similarity). It binds to a Tax-responsive enhancer element in the long terminal repeat of HTLV-I. Regulates the induction of DDIT3/CHOP and asparagine synthetase (ASNS) in response to endoplasmic reticulum (ER) stress. In concert with DDIT3/CHOP, activates the transcription of TRIB3 and promotes ER stress-induced neuronal apoptosis by regulating the transcriptional induction of BBC3/PUMA. Activates transcription of SIRT4. Regulates the circadian expression of the core clock component PER2 and the serotonin transporter SLC6A4. Binds in a circadian time-dependent manner to the cAMP response elements (CRE) in the SLC6A4 and PER2 promoters and periodically activates the transcription of these genes. During ER stress response, activates the transcription of NLRP1, possibly in concert with other factors.

PTMs:

Ubiquitinated by SCF(BTRC) in response to mTORC1 signal, followed by proteasomal degradation and leading to down-regulate expression of SIRT4.

Phosphorylated by NEK6 (By similarity). Phosphorylated on the betaTrCP degron motif at Ser-219, followed by phosphorylation at Thr-213, Ser-224, Ser-231, Ser-235 and Ser-248, promoting interaction with BTRC and ubiquitination. Phosphorylation is promoted by mTORC1 (By similarity).

Phosphorylated by NEK6.

Subcellular Location:

Cytoplasm. Cell membrane. Nucleus. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome.
Note: Colocalizes with GABBR1 in hippocampal neuron dendritic membranes (By similarity). Colocalizes with NEK6 at the centrosome (PubMed:20873783).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

Binds DNA as a homodimer and as a heterodimer. Interacts with CEBPB and binds DNA as a heterodimer with CEBPB. Interacts (via its leucine zipper domain) with GABBR1 and GABBR2 (via their C-termini). Interacts (via its DNA binding domain) with FOXO1 (C-terminal half); the interaction occurs in osteoblasts and regulates glucose homeostasis through suppression of beta-cell proliferation and a decrease in insulin production. Interacts with SATB2; the interaction results in enhanced DNA binding and transactivation by these transcription factors (By similarity). Interacts with CEP290 (via an N-terminal region). Interacts with NEK6, DAPK2 (isoform 2) and ZIPK/DAPK3. Forms a heterodimer with TXLNG in osteoblasts. Interacts with DDIT3/CHOP. Interacts with ABRAXAS2.

Family&Domains:

The BetaTrCP degron motif promotes binding to BTRC when phosphorylated.

Belongs to the bZIP family.

References

1). A marine-derived small molecule induces immunogenic cell death against triple-negative breast cancer through ER stress-CHOP pathway. International Journal of Biological Sciences (PubMed: 35541893) [IF=9.2]

Application: WB    Species: human    Sample: MDA-MB-231 cells

Figure 2. | MHO7 induced ER stress and cell cycle arrest in TNBC cells.(D) The expression of BiP/p-PERK/p-eIF2α/ATF4/CHOP were measured by western blot under MHO7 treatment in MDA-MB-231 cells.
2). Icariside II enhances cisplatin-induced apoptosis by promoting endoplasmic reticulum stress signalling in non-small cell lung cancer cells. International Journal of Biological Sciences (PubMed: 35342361) [IF=9.2]

Application: WB    Species: mouse    Sample: A549/DDP cells

Figure 5.| IS enhances cisplatin-induced apoptosis by promoting ER stress in A549/DDP cells.(H) IS promotes the expression of the proapoptotic transcription factor CHOP. Cells were harvested after 24 h of treatment. Data were from at least three independent experiments. 4-PBA: 4-phyenylbutyric acid.
3). 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.
4). Discontinuation of PTH therapy amplifies bone loss by increasing oxidative stress: An event ameliorated by sequential IL-17 neutralizing antibody therapy. Biomedicine & Pharmacotherapy (PubMed: 34839260) [IF=7.5]
5). Honokiol induces paraptosis-like cell death of acute promyelocytic leukemia via mTOR & MAPK signaling pathways activation. Apoptosis (PubMed: 33550458) [IF=7.2]

Application: IF/ICC    Species: human    Sample: NB4 cells

Fig. 4 | HNK can stimulate the mitochondrial damage and endoplasmic reticulum stress. The NB4 cells were incubated with 0, 10, 20,and 30 μM HNK for 24 h.g The fuorescence intensity of endoplasmic reticulum stress protein ATF4 (green) was observed by immunofuorescence.

Application: IF/ICC    Species: Human    Sample: NB4 cells

Fig. 4 HNK can stimulate the mitochondrial damage and endoplasmic reticulum stress. The NB4 cells were incubated with 0, 10, 20, and 30 μM HNK for 24 h. a A fluorescent probe DCFH-DA detected the expression level of ROS in NB4 cells. 0.1 mg/mL Rosup was used as a positive control. b Measured the relative fluorescence intensity of each group of samples using a microplate reader. c Mitochondrial membrane potential was analyzed by JC-1. 0.1 mM CCCP was used as a positive control. d A specific endoplasmic reticulum fluorescent probe (red) was used to observe the formation of endoplasmic reticulum vacuoles (yellow allows) after 30 μM HNK treatment. e, f The endoplasmic reticulum stress protein BiP and CHOP were analyzed by western blotting. g The fluorescence intensity of endoplasmic reticulum stress protein ATF4 (green) was observed by immunofluorescence. h Real-Time PCR was used to measure the mRNA expression of BiP, CHOP, and ATF4. The results are expressed as mean ± SD (n = 3). Compared with the solvent group, *P < 0.05, **P < 0.01, and ***P < 0.001 (Color figure online)
6). Mitofusin2 Ameliorated Endoplasmic Reticulum Stress and Mitochondrial Reactive Oxygen Species Through Maintaining Mitochondria-Associated Endoplasmic Reticulum Membrane Integrity in Cisplatin-Induced Acute Kidney Injury. Antioxidants & Redox Signaling (PubMed: 37053105) [IF=6.6]
7). Growth differentiation factor-15 overexpression promotes cell proliferation and predicts poor prognosis in cerebral lower-grade gliomas correlated with hypoxia and glycolysis signature. Life Sciences (PubMed: 35588865) [IF=6.1]
8). 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 5 | PA treatment attenuated HFD-induced VLDLR expression in rats. (A) Representative immunoreactive bands of eIF2α, p-eIF2α, ATF4, and VLDLR
9). ATP citrate lyase inhibitor triggers endoplasmic reticulum stress to induce hepatocellular carcinoma cell apoptosis via p‐eIF2α/ATF4/CHOP axis. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (PubMed: 33393219) [IF=5.3]

Application: WB    Species: Human    Sample: HepG2 cells

FIGURE 5 ACLY inhibitor triggers ER stress and activates p‐eIF2α/ATF4/CHOP axis in vitro. Western blot analysis of (A) ER stress‐related proteins (p‐eIF2α, eIF2α, ATF4 and CHOP) and (B) UPR signal transduction molecules (p‐PERK, PERK, p‐IRE1α, IRE1α and sXBP1) in HepG2 cells after administration of BMS‐303141. ATF4p‐eIF2α, eIF2α were activated 3 h post‐treatment; CHOP was activated 8 h post‐treatment. (* P < .05, ** P < .01 and *** P < .001, compared with control group) (C) Western blot analysis of protein expression after ATF4 knockdown. (D) Annexin V‐FITC/PI double staining was performed to determine the apoptosis rate of HepG2 cells after ATF4 knockdown via flow cytometry. (* P < .05, ** P < .01 and *** P < .001, compared with con siRNA group). All experiments were repeated 3 times
10). Protective autophagy alleviates neurotoxin-gelsenicine induced apoptosis through PERK signaling pathway in Neuro-2a cells. TOXICOLOGY (PubMed: 35588915) [IF=4.5]
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