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Key Product Details

Validated by

Independent Antibodies

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse

Applications

Validated:

Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Knockdown Validated, Knockout Validated, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

1 mg/ml

Product Specifications

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

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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.

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.

Scientific Data Images for 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 license.
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.

Applications for IRE1 alpha Antibody

Application
Recommended Usage

Immunocytochemistry/Immunofluorescence

1:100 - 1:250

Immunohistochemistry

1:250 - 1:500

Immunohistochemistry-Paraffin

1:250 - 1:500

Knockout Validated

reported in scientific literature (PMID 31159306)

Western Blot

1:1000 - 1:2000
Application Notes
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. 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.

Reviewed Applications

Read 2 reviews rated 4.5 using NB100-2324 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Glycine, 0.15M NaCl

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: IRE1 alpha

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

Long Name

Serine/threonine-protein kinase/endoribonuclease IRE1

Alternate Names

ERN1, hIRE1p, IRE1, Ire1-alpha

Gene Symbol

ERN1

Additional IRE1 alpha Products

Product Documents for IRE1 alpha Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for IRE1 alpha Antibody

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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