IRE1 alpha [p Ser724] Antibody

Images

 
Immunohistochemistry: IRE1 alpha [p Ser724] Antibody [NB100-2323] - P-Ire1a activation correlates with WFS1 cell expression. Cryosections of retina from 12 month old Wfs1+/+ and Wfs1-/- mouse were immunostained with ...read more
Western Blot: IRE1 alpha [p Ser724] Antibody [NB100-2323] - 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 ...read more
Western Blot: IRE1 alpha [p Ser724] Antibody [NB100-2323] - 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 ...read more
Western Blot: IRE1 alpha [p Ser724] Antibody [NB100-2323] - 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 ...read more
Biological Strategies: Western Blot: IRE1 alpha [p Ser724] Antibody [NB100-2323] - Analysis of anti-IRE1 alpha (pSer724) using Lot AN and AO of NB100-2323. HeLa cells were treated (+) or untreated (-) with 10 mM ...read more
Western Blot: IRE1 alpha [p Ser724] Antibody [NB100-2323] - Regulation of the unfolded protein response by KLF15. Western analysis of UPR activity in WT versus KLF15-/- primary hepatocytes. Hepatocytes were isolated ...read more
Biological Strategies: Western Blot: IRE1 alpha [p Ser724] Antibody [NB100-2323] - Detection in Min6 cells which were treated with different concentrations of glucose for 3 hours prior to lysates preparation.
Immunohistochemistry-Paraffin: IRE1 alpha [p Ser724] Antibody [NB100-2323] - IRE1 (pS724) was detected in immersion fixed paraffin-embedded sections of human spleen using Rabbit Anti-Human IRE1 (pS724) polyclonal ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Dr, Gt, Ma, Pm, Pm, RbSpecies Glossary
Applications WB, ChIP, ELISA, IB, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, IP, KD
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml
Validated by:
   

Biological Strategies

     

IRE1 alpha [p Ser724] Antibody Summary

Description
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.
Immunogen
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]
Modification
p Ser724
Localization
Endoplasmic reticulum
Specificity
NB100-2323 IRE1 alpha [p Ser724] Antibody detects IRE-1 alpha when phosphorylated at Ser724 residue.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
ERN1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:500-1:1000
  • Chromatin Immunoprecipitation
  • ELISA 1:100 - 1:2000
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:10 - 1:500
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Frozen 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:10 - 1:500
  • Immunoprecipitation 1:10 - 1:500
  • In vitro assay
  • Knockdown Validated
Application Notes
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).
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
IRE1 alpha Antibody
Human Pancreas Whole Tissue Lysate (Adult Whole Tumor)
Human Epididymis Whole Tissue Lysate (Adult Whole Normal)
IRE1 alpha Antibody Pack
Control Peptide
IRE1 alpha Protein (NB100-2323PEP)
Reviewed Applications
Read 6 Reviews rated 4.5
using
NB100-2323 in the following applications:

Publications
Read Publications using
NB100-2323 in the following applications:

Reactivity Notes

Drosophila reactivity reported in scientific literature (PMID: 31641108). Goat reactivity reported in scientific literature (PMID: 29046053).

Packaging, Storage & Formulations

Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.025% Sodium Azide
Concentration
1.0 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 [p Ser724] 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
  • ire1 alpha p Ser724
  • ire1 alpha phosphorylation
  • IRE1a
  • ire1-alpha
  • IRE1MGC163279
  • IRE1P
  • MGC163277
  • phospho ire1 alpha
  • phospho-ire1 alpha
  • phospho-ire1a
  • p-ire1 alpha
  • 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.

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Species: Hu, Mu, Rt, Po, Dr, Gt, Ma, Pm, Pm, Rb
Applications: WB, ChIP, ELISA, IB, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, IP, KD

Publications for IRE1 alpha Antibody (NB100-2323)(210)

We have publications tested in 7 confirmed species: Human, Mouse, Rat, Goat, Mammal, Primate, Rabbit.

We have publications tested in 8 applications: ELISA, ICC/IF, IHC, IHC-Fr, IHC-P, IP, Knockdown Validated, WB.


Filter By Application
ELISA
(2)
ICC/IF
(7)
IHC
(7)
IHC-Fr
(1)
IHC-P
(7)
IP
(2)
Knockdown Validated
(1)
WB
(153)
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Human
(74)
Mouse
(77)
Rat
(23)
Goat
(1)
Mammal
(1)
Primate
(1)
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(1)
All Species
Showing Publications 1 - 10 of 210. Show All 210 Publications.
Publications using NB100-2323 Applications Species
Li C, Deng M, Gao Z et al. Effects of compound K, a metabolite of ginsenosides, on memory and cognitive dysfunction in db/db mice involves inhibition of ER stress and NLRP3 inflammasome pathway Food Funct. Apr 13 2020 [PMID: 32374299] (WB) WB
Bellini L, Strub T, Habel N et al. Endoplasmic reticulum stress mediates resistance to BCL-2 inhibitor in uveal melanoma cells Cell Death Discov Apr 17 2020 [PMID: 32337074]
Liu D, Zhang P, Zhou J et al. TNFAIP3 Interacting Protein 3 Overexpression Suppresses Nonalcoholic Steatohepatitis by Blocking TAK1 Activation Cell Metab. Apr 7 2020 [PMID: 32268115]
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) WB Human
Hinte F, van Anken E, Tirosh B, Brune W Repression of viral gene expression and replication by the unfolded protein response effector XBP1u Elife Feb 17 2020 [PMID: 32065579] (WB, Mouse) WB Mouse
Liu XQ, Deng YX, Dai Z et al. Sodium tanshinone IIA sulfonate protects against A alpha 1-42-induced cellular toxicity by modulating A beta-degrading enzymes in HT22 cells Int. J. Biol. Macromol. Feb 6 2020 [PMID: 32035959] (WB, Human) WB Human
Woo M, Choi HI, Park SH et al. The unc-51 like autophagy activating kinase 1-autophagy related 13 complex has distinct functions in tunicamycin-treated cells Biochem. Biophys. Res. Commun. Feb 5 2020 [PMID: 32035621]
Maeyashiki C, Melhem H, Hering L et al. Activation of pH-Sensing Receptor OGR1 (GPR68) Induces ER Stress Via the IRE1 alpha/JNK Pathway in an Intestinal Epithelial Cell Model Sci Rep Jan 29 2020 [PMID: 31996710] (WB, Human) 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) WB Rat
Yoshikawa Y, Sugimoto K, Ochiai Y, Ohashi N Intracellular proliferation of Anaplasma phagocytophilum is promoted via modulation of endoplasmic reticulum stress signaling in host cells Microbiol. Immunol. Jan 7 2020 [PMID: 31909489]
Show All 210 Publications.

Reviews for IRE1 alpha Antibody (NB100-2323) (6) 4.56

Average Rating: 4.5
(Based on 6 reviews)
We have 6 reviews tested in 1 species: Mouse.

Reviews using NB100-2323:
Filter by Applications
WB
(5)
ICC
(1)
All Applications
Filter by Species
Mouse
(6)
All Species
Images Ratings Applications Species Date Details
Western Blot IRE1 alpha NB100-2323
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reviewed by:
Anonymous
WB Mouse 10/22/2019
View

Summary

ApplicationWestern Blot
Sample Testedmacrophage whole cell lysate
SpeciesMouse
LotAW

Comments

Comments1:1000 dilution
Immunocytochemistry IRE1 alpha NB100-2323
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reviewed by:
Anonymous
ICC Mouse 11/19/2018
View

Summary

ApplicationImmunocytochemistry
Sample Testedmouse vascular smooth muscle cell
SpeciesMouse
LotAI-1

Comments

Comments0.1 µM Thapsigargin for 2 hours
Fixation: 4%PFA/PBS for 10 minutes RT
Permeabilization: 50 µg/ml digitonin PBS for 10 minutes RT,
Blocking: 0.1% gelatin/PBS for 30minutes RT
primary antibody for 1 hour RT
secondary anti rabbit IgG Alexa 488 1:200 against p-IRE1 (ser724) for 1 hour RT
Western Blot IRE1 alpha NB100-2323
Enlarge
reviewed by:
Insu Kwon
WB Mouse 04/03/2017
View

Summary

ApplicationWestern Blot
Sample TestedMouse skeletal muscle
SpeciesMouse
LotAO-1

Comments

Comments C57BL/6 Mice
Western Blot IRE1 alpha NB100-2323
Enlarge
reviewed by:
Kihong Lim
WB Mouse 03/15/2017
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysates: Hamster, Mouse, Human, Monkey
SpeciesMouse
LotAO-1

Comments

Comments6132 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:
Anonymous
WB Mouse 12/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 22951927
SpeciesMouse

Blocking

Blocking DetailsSee PMID 22951927

Details

Detection NotesSee PMID 22951927

Comments

CommentsPublished in PMID: 22951927
Western Blot IRE1 alpha NB100-2323
Enlarge
reviewed by:
Anonymous
WB Mouse 05/27/2009
View

Summary

ApplicationWestern Blot
Sample TestedMouse lens tissue, Sample Amount: 40ug
SpeciesMouse
LotI

Blocking

Blocking DetailsBlocking Buffer: 5% TBS+BSA, Blocking Time: overnight, Blocking Temp: 4 degrees Celsius

Primary Anitbody

Dilution RatioDilution Ratio: 1/1000, Incubation Dilution Buffer: TBS+BSA, Incubation Time: 2 hours, Incubation Temp: 24 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: ANTI-RABBIT (CALBIOCHEM), Secondary Ab Dilution Ratio: 1/20000

Details

Detection NotesDetection Method: Chemiluminescence, Exposure Time: 5 minutes, Specific Bands: 110 kDa

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

FAQs for IRE1 alpha Antibody (NB100-2323). (Showing 1 - 4 of 4 FAQs).

  1. 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.
  2. What is the size of the western blot band?
    • The weight observed in our in-house testing is typically 100-110kDa
  3. 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.
  4. 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 at https://www.novusbio.com/support/quality-assurance.html

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 NB100-2323AF405
Alexa Fluor 488 NB100-2323AF488
Alexa Fluor 532 NB100-2323AF532
Alexa Fluor 647 NB100-2323AF647
Alexa Fluor 700 NB100-2323AF700
Biotin NB100-2323B
DyLight 350 NB100-2323UV
DyLight 405 NB100-2323V
DyLight 488 NB100-2323G
DyLight 550 NB100-2323R
DyLight 594 NB100-2323DL594
DyLight 650 NB100-2323C
DyLight 680 NB100-2323FR
DyLight 755 NB100-2323IR
FITC NB100-2323F
HRP NB100-2323H
Janelia Fluor 549 NB100-2323JF549
Janelia Fluor 646 NB100-2323JF646

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Blogs on IRE1 alpha.

Analysis of Total & pSer724 IRE1 alpha, the Sensor of ER Stress
Inositol-requiring protein 1/IRE1 alpha (also called Endoplasmic Reticulum to Nucleus Signaling 1/ERN1; predicted mol wt 110 kDa) is a serine-threonine protein kinase/endoribonuclease which plays a highly critical role in unfolded protein response/U...  Read full blog post.

IRE1 alpha dependent apoptotic-signaling pathway
Despite in depth characterization of the role of IRE1 alpha (inositol-requiring enzyme 1 alpha) in activating the unfolded protein response (UPR) in the ER - little is known about the molecular mechanisms by which this ER protein has shown to reg...  Read full blog post.

IRE1 alpha (inositol-requiring enzyme 1 alpha)
The unfolded protein response (UPR) is a eukaryotic cell process that addresses ER stress. UPR is initiated by three ER-localized sensors: PKR-like ER kinase (PERK), activating transcription factor 6 (ATF6), and inositol-requiring enzyme 1 alpha (...  Read full blog post.

IRE1 alpha and ER Stress: Keys to Disease Progression Pathways
Inositol Requiring Enzyme 1 alpha (IRE1 alpha) is a transmembrane-RNase with an endoplasmic reticulum (ER) luminal sensor domain and cytosolic kinase and ribonuclease domains. IRE1 also plays a central role in the ER stress response (1). In a recent s...  Read full blog post.

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Recent Reviews

4.5
5
3
4
3
3
0
2
0
1
0

Anonymous
10/22/2019
Application: WB
Species: Mouse

Anonymous
11/19/2018
Application: ICC
Species: Mouse

Insu Kwon
04/03/2017
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol ERN1