Product: Phospho-IRE1 (Ser724) Antibody
Catalog: AF7150
Description: Rabbit polyclonal antibody to Phospho-IRE1 (Ser724)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 110kDa; 110kD(Calculated).
Uniprot: O75460
RRID: AB_2843590

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 100ul $350 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 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.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(90%)
Clonality:
Polyclonal
Specificity:
Phospho-IRE1 (Ser724) Antibody detects endogenous levels of IRE1 only when phosphorylated at phospho S724.
RRID:
AB_2843590
Cite Format: Affinity Biosciences Cat# AF7150, RRID:AB_2843590.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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

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; 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;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O75460 ERN1_HUMAN:

Ubiquitously expressed. High levels observed in pancreatic tissue.

Sequence:
MPARRLLLLLTLLLPGLGIFGSTSTVTLPETLLFVSTLDGSLHAVSKRTGSIKWTLKEDPVLQVPTHVEEPAFLPDPNDGSLYTLGSKNNEGLTKLPFTIPELVQASPCRSSDGILYMGKKQDIWYVIDLLTGEKQQTLSSAFADSLCPSTSLLYLGRTEYTITMYDTKTRELRWNATYFDYAASLPEDDVDYKMSHFVSNGDGLVVTVDSESGDVLWIQNYASPVVAFYVWQREGLRKVMHINVAVETLRYLTFMSGEVGRITKWKYPFPKETEAKSKLTPTLYVGKYSTSLYASPSMVHEGVAVVPRGSTLPLLEGPQTDGVTIGDKGECVITPSTDVKFDPGLKSKNKLNYLRNYWLLIGHHETPLSASTKMLERFPNNLPKHRENVIPADSEKKSFEEVINLVDQTSENAPTTVSRDVEEKPAHAPARPEAPVDSMLKDMATIILSTFLLIGWVAFIITYPLSMHQQQQLQHQQFQKELEKIQLLQQQQQQLPFHPPGDTAQDGELLDTSGPYSESSGTSSPSTSPRASNHSLCSGSSASKAGSSPSLEQDDGDEETSVVIVGKISFCPKDVLGHGAEGTIVYRGMFDNRDVAVKRILPECFSFADREVQLLRESDEHPNVIRYFCTEKDRQFQYIAIELCAATLQEYVEQKDFAHLGLEPITLLQQTTSGLAHLHSLNIVHRDLKPHNILISMPNAHGKIKAMISDFGLCKKLAVGRHSFSRRSGVPGTEGWIAPEMLSEDCKENPTYTVDIFSAGCVFYYVISEGSHPFGKSLQRQANILLGACSLDCLHPEKHEDVIARELIEKMIAMDPQKRPSAKHVLKHPFFWSLEKQLQFFQDVSDRIEKESLDGPIVKQLERGGRAVVKMDWRENITVPLQTDLRKFRTYKGGSVRDLLRAMRNKKHHYRELPAEVRETLGSLPDDFVCYFTSRFPHLLAHTYRAMELCSHERLFQPYYFHEPPEPQPPVTPDAL

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
Bovine
100
Sheep
100
Dog
100
Chicken
100
Rabbit
100
Xenopus
90
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O75460 As Substrate

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 As Enzyme

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

Research Backgrounds

Function:

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.

Subcellular Location:

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
Tissue Specificity:

Ubiquitously expressed. High levels observed in pancreatic tissue.

Subunit Structure:

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.

Family&Domains:

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

Research Fields

· 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.

References

1). Melatonin restores endoplasmic reticulum homeostasis to protect injured neurons in a rat model of chronic cervical cord compression. JOURNAL OF PINEAL RESEARCH (PubMed: 36732085) [IF=10.3]

2). ATF4 protects against sorafenib-induced cardiotoxicity by suppressing ferroptosis. BIOMEDICINE & PHARMACOTHERAPY (PubMed: 35724508) [IF=7.5]

3). 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 (PubMed: 37364692) [IF=7.4]

4). Gambogenic acid induces apoptosis and autophagy through ROS‐mediated endoplasmic reticulum stress via JNK pathway in prostate cancer cells. PHYTOTHERAPY RESEARCH (PubMed: 36086867) [IF=7.2]

5). mTOR pathway mediates endoplasmic reticulum stress-induced CD4+ T cell apoptosis in septic mice. APOPTOSIS (PubMed: 35759162) [IF=7.2]

Application: WB    Species: Mice    Sample: CD4+ T cells

Fig. 5 Expression of Akt and IRE1-JNK-related proteins in splenic CD4+ T cells of sepsis mice. Protein levels of Akt, p-Akt, p-IRE1α, JNK, and p-JNK (A-D) in CD4+ T cells were quantified by western blotting and showed as the relative expression values of β-actin, which was used as a loading control to normalize protein levels. For highlighting the activation level of p-Akt and p-JNK in this signaling pathway, the ratio of p-Akt to Akt (p/t Akt) and p-JNK to JNK (p/t JNK) were used for statistics. Data are shown as Mean ± SD (n = 6). Statistically significant differences were determined by one-way analysis of variance (ANOVA) followed by Dunn’s/Tukey’s test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

6). Aqueous Extract of Pepino Leaves Ameliorates Palmitic Acid-Induced Hepatocellular Lipotoxicity via Inhibition of Endoplasmic Reticulum Stress and Apoptosis. Antioxidants (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.

7). PTTG1 alleviates acute alcoholic liver injury by inhibiting endoplasmic reticulum stress‐induced hepatocyte pyroptosis. LIVER INTERNATIONAL (PubMed: 36737842) [IF=6.7]

8). Pterostilbene Prevents Tunicamycin-Induced Intestinal Barrier Damage by Targeting Endoplasmic Reticulum Stress, Oxidative Stress, Autophagy, and Gut Microbiota. Journal of Agricultural and Food Chemistry (PubMed: 36225099) [IF=6.1]

9). Endoplasmic Reticulum Stress Contributes to Copper-Induced Pyroptosis via Regulating the IRE1α-XBP1 Pathway in Pig Jejunal Epithelial Cells. Journal of Agricultural and Food Chemistry (PubMed: 35075900) [IF=6.1]

10). Sesamol supplementation alleviates nonalcoholic steatohepatitis and atherosclerosis in high-fat, high carbohydrate and high-cholesterol diet-fed rats. Food & Function (PubMed: 34606548) [IF=6.1]

Application: WB    Species: Rat    Sample:

Fig. 5 Sesamol regulated the hepatic ERS-IRE1 signaling pathway and NLRP3 expression in HF-HCC diet-fed rats. (A) Representative images of the western blot analyses from hepatic Bip, IRE1, phospho-IRE1, and NLRP3 expression. (B) The semi-quantitative analysis of Bip, and (C and D) representative images of the IHC analyses from hepatic phospho-IRE1 and NLRP3 expression (samples from 3 rats in each group were analyzed). Values are expressed as means ± SEM (n = 3); ###P < 0.001, versus control; ***P < 0.001, versus HF-HCC.

Application: IHC    Species: Rat    Sample:

Fig. 5 Sesamol regulated the hepatic ERS-IRE1 signaling pathway and NLRP3 expression in HF-HCC diet-fed rats. (A) Representative images of the western blot analyses from hepatic Bip, IRE1, phospho-IRE1, and NLRP3 expression. (B) The semi-quantitative analysis of Bip, and (C and D) representative images of the IHC analyses from hepatic phospho-IRE1 and NLRP3 expression (samples from 3 rats in each group were analyzed). Values are expressed as means ± SEM (n = 3); ###P < 0.001, versus control; ***P < 0.001, versus HF-HCC.

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