IRE1 alpha Antibody

Immunohistochemistry: IRE1 alpha Antibody [NB100-2324] - XBP1 and TG2 deficiency promote increased IRE1a protein levels in cultured VSMCs and in neointima in situ. (D) Xbp1WT and Xbp1 delta WT and (E) Tgm2 WT and Tgm2 delta carotid artery sections were analyzed by IHC for total IRE1a. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0212235) licensed under a CC-BY licence.

Western Blot: IRE1 alpha Antibody [NB100-2324] - Total IRE1 Alpha protein in lysates from Min6 cells which were transfected with GFP-siRNA or Ire1-siRNA. Theoretical molecular weight: 110 kDa.

Western Blot: IRE1 alpha Antibody [NB100-2324] - ALOX15 knockout blocks alcohol-induced up-regulation of IRE1a in mouse liver. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: IRE1 alpha Antibody [NB100-2324] - IRE-1 alpha antibody was tested in HeLa cells at 1:200 against Dylight 488 (Green). Alpha tubulin and nuclei were counterstained against Dylight 568 (Red) and DAPI (Blue).

Immunohistochemistry-Paraffin: IRE1 alpha Antibody [NB100-2324] - Analysis of a formalin fixed mouse colon tissue section using total IRE1 alpha antibody (NB100-2324) at 1:300. Note a strong cytoplasmic signal in various cells of colonic villi (columnar epithelial and goblet cells) and the muscular layer. A few cells, potentially with active UPR, showed nuclear positivity also while the signal was negligible in cells at base of crypts and lamina propria.

Immunohistochemistry-Paraffin: IRE1 alpha Antibody [NB100-2324] - Analysis of a formalin fixed mouse colon tissue section using total IRE1 alpha antibody (NB100-2324) at 1:300. Note a strong cytoplasmic signal in various cells of colonic villi (columnar epithelial and goblet cells) and the muscular layer. Some cells, most likely with active UPR, depicted nuclear positivity also while the signal was negligible in crypts cells and lamina propria.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC, IHC-P, KD
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

IRE1 alpha Antibody Summary

• Description: NB100-2324 is useful for the detection of endogenous total IRE1 Alpha protein. For detecting phospho-IRE1 Alpha (Ser-724), we suggest NB100-2323 and it is recommended to normalize phospho-IRE1 alpha band intensity/immunoreactivity with total-IRE1 alpha using NB100-2324 or NB110-59971.
• Immunogen: This IRE1 alpha antibody was raised against a synthetic peptide within the human IRE1 alpha protein (within residues 700-800). [Swiss-Prot #O75460]
• Localization: Endoplasmic reticulum
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ERN1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000 - 1:2000
  • Immunocytochemistry/Immunofluorescence 1:100 - 1:250
  • Immunohistochemistry 1:250 - 1:500
  • Immunohistochemistry-Paraffin 1:250 - 1:500
  • Knockdown Validated
• Application Notes: In Western blot a band at ~110 kDa is observed. In ICC/IF endoplasmic reticulum staining was observed in HeLa cells. 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. Knockout Validated reported in scientific literature (PMID: 31159306).
• Theoretical MW: 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.
• Control:

  Human Pancreas Whole Tissue Lysate (Adult Whole Normal)
  Human Tonsil Whole Tissue Lysate (Adult Whole Normal)
  IRE1 alpha Knockout HeLa Cell Lysate

• Control Peptide:

  IRE1 alpha Protein (NB100-2324PEP)

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine, 0.15M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Notes

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.

Alternate Names for IRE1 alpha Antibody

• EC 2.7.11
• endoplasmic reticulum to nucleus signaling 1
• Endoplasmic reticulum-to-nucleus signaling 1
• ER to nucleus signalling 1
• FLJ30999
• hIRE1p
• inositol-requiring 1
• Inositol-requiring protein 1
• IRE 1a
• IRE1a
• ire1-alpha
• IRE1MGC163279
• IRE1P
• MGC163277
• protein kinase/endoribonuclease
• serine/threonine-protein kinase/endoribonuclease IRE1

Background

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.

References

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

Limitations

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.

Publications for IRE1 alpha Antibody (NB100-2324)(34)

Publications using NB100-2324

• Ding R, Ou W, Chen C et al. Endoplasmic reticulum stress and oxidative stress contribute to neuronal pyroptosis caused by cerebral venous sinus thrombosis in rats: involvement of TXNIP/peroxynitrite-NLRP3 inflammasome activation Neurochem. Int. Sep 24 2020 [PMID: 32980492] (WB, Rat)
• Srinivasan MP, Bhopale KK, Caracheo AA et al. Activation of AMP-activated protein kinase attenuates ethanol-induced ER/oxidative stress and lipid phenotype in human pancreatic acinar cells Biochem Pharmacol Oct 20 2020 [PMID: 32717227] (WB, Human, Rat)
• Cicalese S, Okuno K, Elliott KJ et al. 78 kDa Glucose-Regulated Protein Attenuates Protein Aggregation and Monocyte Adhesion Induced by Angiotensin II in Vascular Cells Int J Mol Sci Jul 15 2020 [PMID: 32679678] (WB, Rat)
• Garufi A, Federici G, Gilardini Montani Ms et Al. Interplay between Endoplasmic Reticulum (ER) Stress and Autophagy Induces Mutant p53H273 Degradation Biomolecules Mar 3 2020 [PMID: 32138264] (WB, Human)
• Han L, Bittner S, Dong D et al. Molecular changes in hepatic metabolism in ZDSD rats-A new polygenic rodent model of obesity, metabolic syndrome, and diabetes Biochim Biophys Acta Mol Basis Dis Jan 24 2020 [PMID: 31987840] (WB, Rat)
• Schmit K, Chen JW, Ayama-Canden S et al. Characterization of the role of TMEM45A in cancer cell sensitivity to cisplatin Cell Death Dis Dec 4 2019 [PMID: 31801939] (WB, Human)
• Gu R, Huang T, Xiao J et al. The IRE1 alpha Arm of UPR Regulates Muscle Cells Immune Characters by Restraining p38 MAPK Activation Front Physiol 2019 Sep 19 [PMID: 31607947] (WB, Mouse)
• Srinivasan MP, Bhopale KK, Amer SM et al. Linking Dysregulated AMPK Signaling and ER Stress in Ethanol-Induced Liver Injury in Hepatic Alcohol Dehydrogenase Deficient Deer Mice Biomolecules Oct 2 2019 [PMID: 31581705] (WB, Mouse)
• Xie W, Xie J, Vince R, More SS Guanabenz Attenuates Acetaminophen-Induced Liver Toxicity and Synergizes Analgesia in Mice Chem. Res. Toxicol. Sep 30 2019 [PMID: 31524377]
• Kaphalia L, Srinivasan MP, Kakumanu RD et al. Ethanol exposure impairs AMPK signaling and phagocytosis in human alveolar macrophages: Role of ethanol metabolism Alcohol. Clin. Exp. Res. Jun 18 2019 [PMID: 31211863]
• Inoue T, Yamakage H, Tanaka M et al. Oxytocin Suppresses Inflammatory Responses Induced by Lipopolysaccharide through Inhibition of the eIF-2-ATF4 Pathway in Mouse Microglia Cells. May 31 2019 [PMID: 31159306] (WB, Knockdown Validated, Mouse)
• Serrano, RL;Yu, W;Graham, RM;Bryan, RL;Terkeltaub, R; A vascular smooth muscle cell X-box binding protein 1 and transglutaminase 2 regulatory circuit limits neointimal hyperplasia PLoS ONE Apr 3 2019 [PMID: 30943188] (IHC-P, Mouse)
• Hsu YH, Chuang HC, Lee YH et al. Traffic-related particulate matter exposure induces nephrotoxicity in vitro and in vivo Free Radic. Biol. Med. Mar 14 2019 [PMID: 30878646] (IHC-P, Rat)
• Lee MR, Bae SJ, Kim JE et al. Inhibition of endoplasmic reticulum stress in high-fat-diet-induced obese C57BL/6 mice: Efficacy of a novel extract from mulberry (Morus alba) leaves fermented with Cordyceps militaris. Lab Anim Res Dec 1 2018 [PMID: 30671117] (WB, Mouse)
• Brenton-Romero R, Weisbrod R, Feng B et all. Liraglutide Treatment Reduces Endothelial Endoplasmic Reticulum Stress and Insulin Resistance in Patients With Diabetes Mellitus. J Am Heart Assoc Sep 18 2018 [PMID: 30371206] (ICC/IF, Human)
• Cho YM, Kim DH, Lee KH et al. The IRE1a-XBP1s pathway promotes insulin-stimulated glucose uptake in adipocytes by increasing PPAR gamma activity Exp. Mol. Med. Aug 15 2018 [PMID: 30111834] (WB, Mouse)
• Yang F, Wang S, Liu Y et al. IRE1a aggravates ischemia reperfusion injury of fatty liver by regulating phenotypic transformation of kupffer cells. Free Radic. Biol. Med. Jun 30 2018 [PMID: 29969718] (WB, Mouse)
• Tripathi M, Zhang CW, Singh BK et al. Hyperhomocysteinemia causes ER stress and impaired autophagy that is reversed by Vitamin B supplementation. Cell Death Dis. Dec 8 2016 [PMID: 27929536] (WB, Mouse)
• Li Y, Zalzala M, Jadhav K et al. Carboxylesterase 2 prevents liver steatosis by modulating lipolysis, ER stress and lipogenesis and is regulated by HNF4alpha. Hepatology. 2016 Jan 25 [PMID: 26806650]
• Kim OK, Jun W, Lee J. Effect of Cudrania tricuspidata and Kaempferol in Endoplasmic Reticulum Stress-Induced Inflammation and Hepatic Insulin Resistance in HepG2 Cells. Nutrients. 2016 Jan 25 [PMID: 26805878] (WB, Human)
• Vannuvel K, Van Steenbrugge M, Demazy C et al. Effects of a Sublethal and Transient Stress of the Endoplasmic Reticulum on the Mitochondrial Population. J. Cell. Physiol. 2015 Dec 17 [PMID: 26680008] (WB, Human)
• Alnasser HA, Guan Q, Zhang F et al. Requirement of clusterin expression for prosurvival autophagy in hypoxic kidney tubular epithelial cells. Am. J. Physiol. Renal Physiol. 2015 Nov 11 [PMID: 26561650] (WB, Mouse)
• Quan X, Wang J, Liang C et al. Melatonin inhibits tunicamycin-induced endoplasmic reticulum stress and insulin resistance in skeletal muscle cells Biochem. Biophys. Res. Commun. 2015 Jun 12 [PMID: 26079879] (WB, Mouse)
  
  Details:
  IRE1 alpha antibody was used for WB on the lysates of C2C12 cells ( a primary line of murine myoblasts) that were serum-starved and treated with 0.5 ug/mL tunicamycin or without in the presence or absence of 100 nM melatonin or 10 uM Luzindole for 16 hours (Fig. 3).
• Clarke JR, Lyra E Silva NM, Figueiredo CP et al. Alzheimer-associated Abeta oligomers impact the central nervous system to induce peripheral metabolic deregulation. EMBO Mol Med 2015 Feb [PMID: 25617315] (WB)
• Ghosh Ak, Garg Sk, Mau T et al. Elevated Endoplasmic Reticulum Stress Response Contributes to Adipose Tissue Inflammation in Aging. J. Gerontol. A Biol. Sci. Med. Sci. 2014 Oct 16 [PMID: 25324219] (WB, Mouse)
• Wu Y, Yang M, Fan J et al. Deficiency of osteoblastic Arl6ip5 impaired osteoblast differentiation and enhanced osteoclastogenesis via disturbance of ER calcium homeostasis and induction of ER stress-mediated apoptosis. Cell Death Dis. 2014 Oct 17 [PMID: 25321471] (WB, Mouse)
• Liu L, Fan Z, Tang Y, Ke Z. The Resveratrol Attenuates Ethanol-Induced Hepatocyte Apoptosis Via Inhibiting ER-Related Caspase-12 Activation and PDE Activity In Vitro. Alcohol Clin Exp Res. 2013 Nov 13 [PMID: 24224909] (WB, Human)
• Zheng M, Zhang Q, Joe Y et al. Carbon Monoxide Releasing Molecules Reverse Leptin Resistance Induced by Endoplasmic Reticulum Stress. Am J Physiol Endocrinol Metab 2013 Feb 12 [PMID: 23403944] (WB, Human, Mouse)
• Arshad M, Ye Z, Gu X et al. RNF13, a RING Finger Protein, Mediates Endoplasmic Reticulum Stress-induced Apoptosis through the Inositol-requiring Enzyme (IRE1alpha)/c-Jun NH2-terminal Kinase Pathway. J Biol Chem 2013 Mar 22 [PMID: 23378536] (WB, Human)
• Kurata M, Yamazaki Y, Kanno Y et al. Anti-apoptotic function of Xbp1 as an IL-3 signaling molecule in hematopoietic cells. Cell Death and Dis. 2011 Feb 10. [PMID: 21368889] (WB, Mouse)
• Luo D, He Y, Zhang H et al. AIP1 is critical in transducing IRE1-mediated endoplasmic reticulum stress response. J Biol Chem;283(18):11905-12. 2008 May 2. [PMID: 18281285] (WB, Mouse)
• Lipson, KL et al. Regulation of insulin biosynthesis in pancreatic beta cells by an endoplasmic reticulum-resident protein kinase IRE1. Cell Metab;4(3):245-54. 2006 Sep. [PMID: 16950141] (WB, Mouse)
• Fonesca, SG et al. WFS1 is a novel component of the unfolded protein response and maintains homeostasis of the endoplasmic reticulum in pancreatic beta-cells. J Biol Chem280(47):39609-15. 2005 Nov 25. [PMID: 16195229]
• Kubota, K et al. Fluoride induces endoplasmic reticulum stress in ameloblasts responsible for dental enamel formation. J Biol Chem;280(24):23194-202. 2005 Jun 17. [PMID: 15849362]

Reviews for IRE1 alpha Antibody (NB100-2324) (2)

Average Rating: 4.5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Mouse.

reviewed by: Anonymous ICC Mouse 11/19/2018

Summary

• Application: Immunocytochemistry
• Sample Tested: mouse vascular smooth muscle cell
• Species: Mouse
• Lot: D-1

reviewed by: Anonymous WB Mouse 03/07/2018

Summary

• Application: Western Blot
• Sample Tested: Liver
• Species: Mouse
• Lot: c1

FAQs for IRE1 alpha Antibody (NB100-2324). (Showing 1 - 2 of 2 FAQs).

1. I have a customer who is interested in antibody NB100-2324 for use with rat samples. It is listed as working with rat and for Western blot applications. Can you tell me which rat tissues were tested in Western blot?
   • Previous productions were reported to work in rat. We have not tested the current lot directly on any rat samples so I do not have data on what tissues, or lysates were effective. We will still guarantee rat as a reactive species, as it is stated on our datasheet.
2. We hope to know which position the antibody NB 100-2324 recognize within the endogenous IRE1 alpha. Thanks. Is the region 700 to 800 within the luminal domain, or transmembrane or cytosolic domain? Is this region highly conserved between Human and mouse?
   • It looks like the 700-800 region is cytoplasmic. The homology in this region is 93%, and this product has already been validated to work in both human and mouse species.https://www.uniprot.org/uniprot/O75460

Bioinformatics

• Gene Symbol: ERN1