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
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.
Genetic Strategies: 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 ...read more
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
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.
A synthetic peptide surrounding the phosphorylated serine 724 of the human IRE1 alpha protein. [Swiss-Prot #O75460]
NB100-2323 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).
Mammal reactivity reported in scientific literature (PMID: 27180817).
Packaging, Storage & Formulations
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
0.025% 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.
Alternate Names for IRE1 alpha [p Ser724] Antibody
Unfolded protein response (UPR) signaling, a mechanism used by eukaryotic cells to cope ER stress. It 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). UPR-responsive genes's transcriptional activation is regulated by ATF6 and IRE1-XBP1 pathways, and UPR serves three important functions: inhibition of protein translation to restore normal cell functions; activation of signaling to increase production of molecular chaperones involved in protein folding; and activation of signaling that leads to targeting of misfolded proteins in ER for ubiquitination and subsequent degradation; or when ER-stress is not relieved, UPR leads to apoptosis. Localized as a single-pass type I membrane protein in ER membrane, IRE1 alpha is ubiquitously expressed with high levels observed in pancreatic tissue. It undergoes autophosphorylation and gets ADP-ribosylated by PARP16 upon ER stress, which increases its kinase as well as endonuclease activities. IRE1 alpha senses unfolded proteins in the ER lumen via its N-terminal domain which leads to enzyme auto-activation and thereafter, the active endoribonuclease domain splices XBP1 mRNA to generate a new C-terminus, converting it into a potent UPR transcriptional activator. IRE1 alpha inactivation in mice has been shown to result in widespread developmental defects, leading to intra-gestational embryonic lethality and ER stress mediated cell death is responsible for several pathologies.
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.
Lu G, Ota A, Ren S et al. PPM1l encodes an inositol requiring-protein 1 (IRE1) specific phosphatase that regulates the functional outcome of the ER stress response. Molecular Metabolism. 2013 Aug (WB, Mouse)
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).
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 at https://www.novusbio.com/support/quality-assurance.html
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