P-Ire1a activation correlates with WFS1 cell expression. Cryosections of retina from 12 month old Wfs1+/+ and Wfs1-/- mouse were immunostained with anti-P-Ire1a antibody (red). DAPI was used for staining of cell nuclei ...read more
Hepatic ER stress markers with cachexia progression. ER stress markers Bip, IRE1a, ATF6p50 and CHOP were examined in the liver of non, pre and severely cachectic mice. (n = 6-8 per group, p < 0.05) Dotted line indicates ...read more
IRF-1 plays a central role in ER stress-mediated modulation of VCAM-1 expression by TGRL. HAEC were conditioned for 4 hr with TNFa (0.3 ng/ml) E: CHOP knockdown decreased TNFa-induced VCAM-1 expression. n=4. **P<0.01 ...read more
Analysis using HRP conjugate of NB100-2323. Detection of phosphorylated IRE-1 alpha using NB100-2323. Lane 1: COS-7 untransfected Lane 2: COS-7 expressing wild-type IRE1 alpha Lane 3: COS-7 expressing kinase-dead IRE1 ...read more
Biological Strategies: Analysis of anti-IRE1 alpha (pSer724) using Lot AN and AO of NB100-2323. HeLa cells were treated (+) or untreated (-) with 10 mM DTT for 60 min to activate the UPR. Total protein was ...read more
Regulation of the unfolded protein response by KLF15. Western analysis of UPR activity in WT versus KLF15-/- primary hepatocytes. Hepatocytes were isolated from standard chow-fed 4-month-old male WT and KLF15 -/- mice. ...read more
HeLa cell lysate, 20 ug. Antibody at 1:1000, 1% skim milk in TBST, overnight incubation at 4C. WB image submitted by a verified customer review.
Biological Strategies: Detection in Min6 cells which were treated with different concentrations of glucose for 3 hours prior to lysates preparation.
IRE1 (pS724) was detected in immersion fixed paraffin-embedded sections of human spleen using Rabbit Anti-Human IRE1 (pS724) polyclonal Antibody (Catalog # NB100-2323) at 1:300 for 1 hour at room temperature followed by ...read more
When detecting phospho-IRE1 Alpha (Ser-724) using NB100-2323, it is recommended to normalize its band intensity/immunoreactivity with total-IRE1 alpha. NB100-2324 and NB110-59971 can be used for the detection of endogenous total IRE1 alpha.
This IRE1 alpha [p Ser724] antibody was raised against a synthetic peptide surrounding the phosphorylated serine 724 of the human IRE1 alpha protein. [Swiss-Prot #O75460]
NB100-2323 IRE1 alpha [p Ser724] Antibody detects IRE-1 alpha when phosphorylated at Ser724 residue.
Immunogen affinity purified
Test in a species/application not listed above to receive a full credit towards a future purchase.
This IRE1 pS724 antibody is useful for WB, ELISA, and IHC-P sections (PMID: 19264902). Use in IHC-Frozen reported in scientific literature (PMID: 24823368). Use in ICC reported in scientific literature (PMID: 26762342). Use in immunoblotting reported in scientific literature (PMID: 24089213). Use In vitro assay reported in scientific literature (PMID: 24327956). Use in chromatin immunoprecipitation reported in scientific literature (PMID: 25225294). Knockdown Validated reported in scientific literature (PMID: 31159306).
110 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Use in Porcine reported in scientific literature (PMID:35492579).Drosophila reactivity reported in scientific literature (PMID: 31641108). Goat reactivity reported in scientific literature (PMID: 29046053). Use in Golden Syrian Hamster reported in scientific literature (PMID:31167774).
Packaging, Storage & Formulations
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
0.02% Sodium Azide
Immunogen affinity purified
Take a look at IRE1 alpha Antibody Sampler Pack [NBP2-50067] if you want to try 25ug aliquots of phospho-IRE1 alpha (Ser724) Antibody [NB100-2323SS] and total IRE1 alpha Antibody [NB100-2324SS] before purchasing 100ug full vials.
Accumulation of misfolded proteins in the endoplasmic reticulum (ER) activates the unfolded protein response (UPR) and upregulates ER molecular chaperones in order to cope with ER stress. UPR is initiated by three ER-localized protein sensors: PERK (PKR-like ER kinase), ATF (activating transcription factor 6), and IRE1 alpha (inositol-requiring enzyme 1 alpha) (1). IRE1 alpha is correlated with X-box binding protein (XBP1) as a potent UPR transcriptional activator (2).
Transcriptional activation of UPR-responsive genes are regulated by the ATF6 and IRE1-XBP1 pathways, which are often regulated by HIFs and contribute to cell survival under ER hypoxic stress (3). UPR signaling can a) inhibit protein translation to restore cell function, b) activate signaling to increase production of molecular chaperones for protein folding, and c) initiate ubiquitination signaling that leads to degradation of misfolded proteins in ER.
IRE1 alpha acts as the sensor of unfolded proteins in the ER. IRE1 alpha not only promotes cell survival but can initiate apoptosis when accumulation of unfolded proteins in the ER causes stress. IRE1 alpha is essential for viability under stress conditions that cause unfolded proteins to accumulate in the ER. IRE1 alpha is a transmembrane protein that has both serine-threonine kinase and endoribonuclease activities and has a theoretical molecular weight of 110 kDa. When detecting phospho-IRE1 alpha, it is recommended to normalize its band intensity with total IRE1 alpha.
1. Zheng, W., Xie, W., Yin, D., Luo, R., Liu, M., & Guo, F. (2019). ATG5 and ATG7 induced autophagy interplays with UPR via PERK signaling. Cell Commun Signal, 17(1), 42. doi:10.1186/s12964-019-0353-3
2. Cho, Y. M., Kim, D. H., Lee, K. H., Jeong, S. W., & Kwon, O. J. (2018). The IRE1alpha-XBP1s pathway promotes insulin-stimulated glucose uptake in adipocytes by increasing PPARgamma activity. Exp Mol Med, 50(8), 102. doi:10.1038/s12276-018-0131-0
3. Xia, Z., Wu, S., Wei, X., Liao, Y., Yi, P., Liu, Y., . . . Liu, J. (2019). Hypoxic ER stress suppresses beta-catenin expression and promotes cooperation between the transcription factors XBP1 and HIF1alpha for cell survival. J Biol Chem, 294(37), 13811-13821. doi:10.1074/jbc.RA119.008353
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Ricci D, Tutton S, Marrocco I et al. The connector between the kinase and RNase domains of IRE1alpha transmits the conformational change that underlies ER stress-induced activation. bioRxiv 2020-01-01 (WB, Human)
Imbernon Piedra M. Efectos centrales de MCH sobre el metabolismo periferico. Thesis 2016-06-01 (IF/IHC, WB)
Kim J, Youn I, Seo E, Kim C Protective effects of Allium macrostemon extract on adipose tissue and liver dysregulation induced by high-fat diet and bisphenol A in C57BL/6 mice Research Square 2023-06-27 (WB, Mouse)
Hong M. Regulation of Human Pancreatic B-cell Proliferation by Localization, Interaction, and Transcription of Endogenously Expressed Cell Cycle Molecules Thesis 2017-01-01 (Human)
Rat placenta lysates, reduced and denatured. 100 ug total protein resolved on 7.5% SDS-acrylamide gel. Transfer onto 0.22 um nitrocellulose membrane. Block with 50% Rockland blocking solution. Incubate membrane overnight at 4C with 1:500 dilution of pIRE1a antibody. Washes with PBS-0.1% tween 20. Incubate for one hour at room temp with 1:10 000 LiCor IRDye 800CW donkey anti-rabbit IgG. Images taken with Odyssey V3.0.
6132 Pro cells (mouse fibrosarcoma) were treated with or without 10 mM DTT for 3 hr and total lysates were prepared. The membrane blot was incubated with 5 ul of primary antibody (IRE1 alpha [p Ser724]) in 10 ml of blocker (2% BSA in TBS containing 0.05% Tween20) for 2hr at RT, followed by incubation with HRP-conjugated secondary antibody (anti-rabbit IgG H+L, from Pierce) for 1hr. The signal was visualized using chemilluminedcent substrate (Pico, from Pierce).
reviewed by: cynthia morimoto
See PMID 22951927
reviewed by: System Admin
Mouse lens tissue, Sample Amount: 40ug
Product General Protocols
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FAQs for IRE1 alpha Antibody (NB100-2323). (Showing 1 - 4 of 4 FAQs).
Can you please tell me the immunogen sequence of this antibody?
The immunogen of NB100-2323 is a synthetic peptide from an amino acids sequence surrounding the phosphorylated serine 724 of the human IRE1 alpha protein (Swiss-Prot #O75460). Unfortunately, I cannot provide any additional detail, as that is proprietary information.
What is the size of the western blot band?
The weight observed in our in-house testing is typically 100-110kDa
Can this antibody detect non-phosphorylated IRE1a?
NB100-2323 is phospho-specific. In all of our testing done with this antibody, we have only detected the phosphorylated IRE1a protein. There has not been any observed cross reactivity with the non-phosphorylated protein.
I wish to use this antibody for cultured cardiomyocytes /Immunofluorescence. I am wondering if this antibody works for Immunocytochemistry/Immunofluorescence for cultured cells? Specifically, we wish to compare IRE1 phosphorylation under stress condition to non-stress condition. Because the nature of IRE1 phosphorylation is part of stress response pathway. I am wondering if the immunofluorescence with your antibody is sensitive enough to distinguish p-IRE1 signal under stress condition to the endogenous stress (no stress inducer treatment) in the cell?
IRE1 p Ser724 antibody (NB100-2323) antibody has been validated in Immunocytochemistry/Immunofluorescence (ICC/IF) and we guarantee it 100% for the applications and species listed on our datasheet. As long as your cultured cardiomyocytes express the protein target that is phosphorylated at Ser 724 amino acid residue, our antibody will detect it. You may learn more about our guaranteee policy here
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