IRE1 Antibody - #DF7709

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製品: IRE1 Antibody
カタログ: DF7709
タンパク質の説明: Rabbit polyclonal antibody to IRE1
アプリケーション: WB IHC IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
分子量: 110 kDa; 110kD(Calculated).
ユニプロット: O75460
RRID: AB_2841178

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MS Report
HPLC Report
MSDS
説明書
COA
プロトコル
WB Handbook
IHC Protocol
IF/ICC Protocol

製品説明

ソース:
Rabbit
アプリケーション:
WB 1:1000-3000, IF/ICC 1:200-1:500, IHC 1:50-1:200
*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.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(86%)
クローナリティ:
Polyclonal
特異性:
IRE1 Antibody detects endogenous levels of total IRE1.
RRID:
AB_2841178
引用形式: Affinity Biosciences Cat# DF7709, RRID:AB_2841178.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
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.
別名:

折りたたみ／展開

Endoplasmic reticulum (ER) to nucleus signalling 1; Endoplasmic reticulum to nucleus signaling 1; Endoplasmic reticulum-to-nucleus signaling 1; Endoribonuclease; ER to nucleus signaling 1; ERN 1; Ern1; ERN1_HUMAN; hIRE 1p; hIRE1p; Inositol requiring 1; Inositol requiring 1, S. cerevisiae, homolog of; Inositol requiring enzyme 1, S. cerevisiae, homolog of; Inositol requiring protein 1; inositol-requiring enzyme 1; Inositol-requiring protein 1; IRE 1; IRE 1a; IRE 1P; Ire1 alpha; Ire1-alpha; IRE1a; Ire1alpha; IRE1P; MGC163277; MGC163279; Protein kinase/endoribonuclease; RGD1559716; Serine/threonine protein kinase/endoribonuclease IRE1;

免疫原

免疫原:
Uniprot:

O75460(ERN1_HUMAN) >>Visit HPA database.

遺伝子(ID):
ERN1(2081)
発現特異性:
O75460 ERN1_HUMAN:

Ubiquitously expressed. High levels observed in pancreatic tissue.

タンパク質配列:
MPARRLLLLLTLLLPGLGIFGSTSTVTLPETLLFVSTLDGSLHAVSKRTGSIKWTLKEDPVLQVPTHVEEPAFLPDPNDGSLYTLGSKNNEGLTKLPFTIPELVQASPCRSSDGILYMGKKQDIWYVIDLLTGEKQQTLSSAFADSLCPSTSLLYLGRTEYTITMYDTKTRELRWNATYFDYAASLPEDDVDYKMSHFVSNGDGLVVTVDSESGDVLWIQNYASPVVAFYVWQREGLRKVMHINVAVETLRYLTFMSGEVGRITKWKYPFPKETEAKSKLTPTLYVGKYSTSLYASPSMVHEGVAVVPRGSTLPLLEGPQTDGVTIGDKGECVITPSTDVKFDPGLKSKNKLNYLRNYWLLIGHHETPLSASTKMLERFPNNLPKHRENVIPADSEKKSFEEVINLVDQTSENAPTTVSRDVEEKPAHAPARPEAPVDSMLKDMATIILSTFLLIGWVAFIITYPLSMHQQQQLQHQQFQKELEKIQLLQQQQQQLPFHPPGDTAQDGELLDTSGPYSESSGTSSPSTSPRASNHSLCSGSSASKAGSSPSLEQDDGDEETSVVIVGKISFCPKDVLGHGAEGTIVYRGMFDNRDVAVKRILPECFSFADREVQLLRESDEHPNVIRYFCTEKDRQFQYIAIELCAATLQEYVEQKDFAHLGLEPITLLQQTTSGLAHLHSLNIVHRDLKPHNILISMPNAHGKIKAMISDFGLCKKLAVGRHSFSRRSGVPGTEGWIAPEMLSEDCKENPTYTVDIFSAGCVFYYVISEGSHPFGKSLQRQANILLGACSLDCLHPEKHEDVIARELIEKMIAMDPQKRPSAKHVLKHPFFWSLEKQLQFFQDVSDRIEKESLDGPIVKQLERGGRAVVKMDWRENITVPLQTDLRKFRTYKGGSVRDLLRAMRNKKHHYRELPAEVRETLGSLPDDFVCYFTSRFPHLLAHTYRAMELCSHERLFQPYYFHEPPEPQPPVTPDAL

種類予測

種類予測:

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
Bovine
100
Sheep
100
Dog
100
Chicken
100
Rabbit
100
Xenopus
86
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O75460 基板として

Site PTM Type Enzyme
T49 Phosphorylation
K88 Ubiquitination
K95 Ubiquitination
T132 Phosphorylation
Y161 Phosphorylation
Y166 Phosphorylation
Y179 Phosphorylation
T283 Phosphorylation
Y289 Phosphorylation
K374 Acetylation
K374 Ubiquitination
K485 Ubiquitination
S533 Phosphorylation
S536 Phosphorylation
S544 Phosphorylation
S548 Phosphorylation
S551 Phosphorylation
Y628 Phosphorylation O75460 (ERN1)
K633 Ubiquitination
K706 Ubiquitination
S724 Phosphorylation O75460 (ERN1)
S726 Phosphorylation
S729 Phosphorylation
K851 Ubiquitination
K860 Ubiquitination
T973 Phosphorylation

PTMs - O75460 酵素として

Substrate Site Source
O75460 (ERN1) Y628 Uniprot
O75460 (ERN1) S724 Uniprot

研究背景

機能:

Serine/threonine-protein kinase and endoribonuclease that acts as a key sensor for the endoplasmic reticulum unfolded protein response (UPR). In unstressed cells, the endoplasmic reticulum luminal domain is maintained in its inactive monomeric state by binding to the endoplasmic reticulum chaperone HSPA5/BiP. Accumulation of misfolded protein in the endoplasmic reticulum causes release of HSPA5/BiP, allowing the luminal domain to homodimerize, promoting autophosphorylation of the kinase domain and subsequent activation of the endoribonuclease activity. The endoribonuclease activity is specific for XBP1 mRNA and excises 26 nucleotides from XBP1 mRNA. The resulting spliced transcript of XBP1 encodes a transcriptional activator protein that up-regulates expression of UPR target genes. Acts as an upstream signal for ER stress-induced GORASP2-mediated unconventional (ER/Golgi-independent) trafficking of CFTR to cell membrane by modulating the expression and localization of SEC16A.

PTMs:

Autophosphorylated following homodimerization. Autophosphorylation promotes activation of the endoribonuclease domain.

ADP-ribosylated by PARP16 upon ER stress, which increases both kinase and endonuclease activities.

細胞の位置付け:

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

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Ubiquitously expressed. High levels observed in pancreatic tissue.

サブユニット構造:

Monomer. Homodimer; disulfide-linked; homodimerization takes place in response to endoplasmic reticulum stress and promotes activation of the kinase and endoribonuclease activities. Dimer formation is driven by hydrophobic interactions within the N-terminal luminal domains and stabilized by disulfide bridges. Interacts (via the luminal region) with DNAJB9/ERdj4; interaction takes place in unstressed cells and promotes recruitment of HSPA5/BiP. Interacts (via the luminal region) with HSPA5/BiP; HSPA5/BiP is a negative regulator of the unfolded protein response (UPR) that prevents homodimerization of ERN1/IRE1 and subsequent activation of the protein. Interacts with PDIA6, a negative regulator of the UPR; the interaction is direct and disrupts homodimerization. Interacts with DAB2IP (via PH domain); the interaction occurs in a endoplasmic reticulum stress-induced dependent manner and is required for subsequent recruitment of TRAF2 to ERN1/IRE1 (By similarity). Interacts with TAOK3 and TRAF2. Interacts with RNF13.

タンパク質ファミリー:

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.

研究領域

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

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

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

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

参考文献

1). Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. NUCLEIC ACIDS RESEARCH, 2020 (PubMed: 32710621) [IF=16.6]

Application: WB    Species: mouse    Sample: liver

Figure 7. Effect of A22 on ameliorating apoptosis, ER stress, inflammation, metabolic syndrome, and fibrogenesis in HF diet-fed mice. (A) Effect of A22 on BCL-2 gene transcription. (B) Effect of A22 on BAX gene transcription. (C) Effect of A22 on expressions of apoptosis-related proteins in liver. The extracted proteins from the liver were immunoblotted with specific antibodies, and quantified based on the loading control of ACTIN. (D) Effect of A22 on ER stress. The UPR proteins (IRE-1, PERK, elF-2 and CHOP) were analyzed by using western Blot. (E) Effect of A22 on expressions of inflammatory factors. (F) Effect of A22 on expressions of fibrogenic proteins.
2). Menaquinone-4 prevents medication-related osteonecrosis of the jaw through the SIRT1 signaling-mediated inhibition of cellular metabolic stresses-induced osteoblast apoptosis. Free Radical Biology and Medicine, 2023 (PubMed: 37364692) [IF=7.1]
3). Aqueous Extract of Pepino Leaves Ameliorates Palmitic Acid-Induced Hepatocellular Lipotoxicity via Inhibition of Endoplasmic Reticulum Stress and Apoptosis. Antioxidants, 2021 (PubMed: 34204987) [IF=7.0]

Application: WB    Species: Human    Sample: HepG2 Cells

Figure 5 AEPL attenuated ER stress while PA exposure. (a) ER stress-related protein levels were analyzed by Western blotting. (b–d) GRP78, phosphorylated PERK, and phosphorylated IRE1αwere normalized to PERK, IRE1α, and β-actin of each sample. (e) SREBP-1 level in each group was normalized to β-actin. All data were presented as mean ± SD of at least three independent experiments. * p < 0.05 vs. control cells; # p < 0.05 vs. PA-treated cells.
4). 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, 2023 (PubMed: 37654182) [IF=5.7]

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

Application: IF/ICC    Species: Human    Sample: HBE135-E6E7 cells

Figure 6 The levels of p-IRE1α, IRE1α and NOD1 expression, and NF-κB activation-nuclear translocation. The cells were transfected with negative control (NC) siRNA, IRE1α siRNA or NOD1 siRNA respectively, or were pretreated with 10 μg/ml C12-iE-DAP (a NOD1 agonist) for 2 h prior to exposure to 100 μg/ml PM2.5 for 24 h. (A) The mRNA expression levels of IRE1α relative to β-actin measured by RT-qPCR. (B) The mRNA levels of NOD1 relative to β-actin. (C) Relative protein expression levels of p-IRE1α, detected by western blot, were depicted as the ratio of each group to β-actin. (D) The expression levels of IRE1α, assayed by the immunofluorescence labeled by CY3, were detected using a fluorescence microscope at x400 magnification. (E) Relative protein expression levels of NOD1, detected using western blot analysis. (F) The expression levels of NOD1, assayed using immunofluorescence labeled with FITC, were detected using a fluorescence microscope at x400 magnification. (G) The intracellular NF-κB activation-nuclear translocation, assayed using a specialized kit, were also detected using a fluorescence microscope at x400 magnification, and the mean gray values were computed using ImageJ software. Data are presented as the mean ± SD (RT-qPCR and western blot with n=3 repeats per group, and immunofluorescence with n=3 different field images per group). *P
5). Pterostilbene Prevents Tunicamycin-Induced Intestinal Barrier Damage by Targeting Endoplasmic Reticulum Stress, Oxidative Stress, Autophagy, and Gut Microbiota. Journal of Agricultural and Food Chemistry, 2022 (PubMed: 36225099) [IF=5.7]
6). Chrysin ameliorates synovitis and fibrosis of osteoarthritic fibroblast-like synoviocytes in rats through PERK/TXNIP/NLRP3 signaling. Frontiers in Pharmacology, 2023 (PubMed: 37021049) [IF=5.6]
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, 2019 (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). Naringin attenuates inflammatory injury to the bovine endometrium by regulating the endoplasmic reticulum stress-PI3K/AKT-autophagy axis. Frontiers in pharmacology, 2024 (PubMed: 39234103) [IF=5.6]

Application: WB    Species: bovine    Sample: bEECs

FIGURE 4. Effects of naringin on LPS-induced ERS. (A) The protein expressions of p-PERK, PERK, pIREα, IREα, and cleaved ATF6 were determined in triplicate by using Western blot analysis. n = 3. (B,C) A confocal image provided by immunofluorescence determined the expression levels of ATF4, XBP1, and ATF6 in bEECs. Scale bars: 50 μm and 10 μm (bottom, n = 2). (D) RT-qPCR analysis of ATF4, CHOP, and GRP78 mRNA expression normalized to the expression of GAPDH. n = 4. The data are presented as the mean ± SEM. Experiments were repeated n times with duplicate biological replicates. *p < 0.05; **p < 0.01; ***p < 0.001.
9). Flavonoids from Hippophae rhamnoides Linn. Revert Doxorubicin-Induced Cardiotoxicity through Inhibition of Mitochondrial Dysfunction in H9c2 Cardiomyoblasts In Vitro. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2023 (PubMed: 36834585) [IF=5.6]
10). 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, 2021 (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
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