Overview

  • Product name
    Anti-IRE1 (phospho S724) antibody
    See all IRE1 primary antibodies
  • Description
    Rabbit polyclonal to IRE1 (phospho S724)
  • Specificity
    ab48187 detects IRE1 alpha, phosphorylated (serine 724) protein.
  • Tested applications
    Suitable for: WB, IHC-P, ELISA, IP, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Does not react with: Monkey
  • Immunogen

    Synthetic peptide corresponding to Human IRE1 (phospho S724).
    Database link: O75460
    (Peptide available as ab110445)

  • Positive control
    • Glucose treated pancreatic islets and insulinoma cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab48187 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/2000. Predicted molecular weight: 110 kDa.

Block with 3-5% BSA.

IHC-P 1/10 - 1/500.
ELISA 1/100 - 1/2000.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 20477942

Target

  • Function
    Senses unfolded proteins in the lumen of the endoplasmic reticulum via its N-terminal domain which leads to enzyme auto-activation. The active endoribonuclease domain splices XBP1 mRNA to generate a new C-terminus, converting it into a potent unfolded-protein response transcriptional activator and triggering growth arrest and apoptosis.
  • Tissue specificity
    Ubiquitously expressed. High levels observed in pancreatic tissue.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
    Contains 1 KEN domain.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Autophosphorylated.
  • Cellular localization
    Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Endoplasmic reticulum (ER) to nucleus signalling 1 antibody
    • Endoplasmic reticulum to nucleus signaling 1 antibody
    • Endoplasmic reticulum-to-nucleus signaling 1 antibody
    • Endoribonuclease antibody
    • ER to nucleus signaling 1 antibody
    • ERN 1 antibody
    • Ern1 antibody
    • ERN1_HUMAN antibody
    • hIRE 1p antibody
    • hIRE1p antibody
    • Inositol requiring 1 antibody
    • Inositol requiring 1, S. cerevisiae, homolog of antibody
    • Inositol requiring enzyme 1, S. cerevisiae, homolog of antibody
    • Inositol requiring protein 1 antibody
    • inositol-requiring enzyme 1 antibody
    • Inositol-requiring protein 1 antibody
    • IRE 1 antibody
    • IRE 1a antibody
    • IRE 1P antibody
    • Ire1 alpha antibody
    • Ire1-alpha antibody
    • IRE1a antibody
    • Ire1alpha antibody
    • IRE1P antibody
    • MGC163277 antibody
    • MGC163279 antibody
    • Protein kinase/endoribonuclease antibody
    • RGD1559716 antibody
    • Serine/threonine protein kinase/endoribonuclease IRE1 antibody
    see all

Images

  • All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 1/2000 dilution

    Lane 1 : HeLa cells 30 nM Calyculin A: AP-buffer
    Lane 2 : HeLa cells 30 nM Calyculin A: Alkaline Phosphatase

    Lysates/proteins at 20 µg per lane.

    Secondary
    infrared (IR)-labelled goat anti-rabbit (green) antibody and IR-labelled goat anti-mouse (red) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 110 kDa

    The blots were produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membranes were blocked for an hour. Membrane 2 was incubated with alkaline phosphatase (AP; 100 U per mL) for one hour, whilst membrane 1 was treated with AP-buffer only, before being  incubated with ab48187 (rabbit anti-IRE1 antibody diluted 1:2000) and loading control ab125247 (mouse anti-GAPDH antibody; diluted 1:10,000) for 24 hours at 4°C. Antibody binding was detected using infrared (IR)-labelled goat anti-rabbit (green) antibody and IR-labelled goat anti-mouse (red) at 1:10,000 dilutions for 1 hour at room temperature before imaging.

  • ab48187 staining IRE1 (phospho S724) in rat testis tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with formaldehyde, permeabilized using 0.1% Triton X-100 and then blocked with 2% BSA for 1 hour at 25°C, followed by incubation with the primary antibody at a 1/200 dilution, for 1 hour at 25°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution.

    See Abreview

  • Serially diluted ab48187 was bound to immobilised Phospho peptide (133861) - or Control peptide (1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed with TMB substrate.

  • All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 1/1000 dilution

    Lane 1 : Min6 cells untreated
    Lane 2 : Min6 cells treated with glucose for 3 hours at 5 mM
    Lane 3 : Min6 cells treated with glucose for 3 hours at 20 mM

    Developed using the ECL technique

    Predicted band size : 110 kDa
  • All lanes : Anti-IRE1 (phospho S724) antibody (ab48187) at 1/2000 dilution

    Lane 1 : Cell lysate prepared from COS-7 Untransfected cells
    Lane 2 : Cell lysate prepared from COS-7 cells expressing wild type IRE1 alpha
    Lane 3 : Cell lysate prepared from COS-7 cells expressing kinase-dead IRE1 alpha


    Predicted band size : 110 kDa

References

This product has been referenced in:
  • Mohammed-Ali Z  et al. Endoplasmic reticulum stress inhibition attenuates hypertensive chronic kidney disease through reduction in proteinuria. Sci Rep 7:41572 (2017). IHC ; Mouse . Read more (PubMed: 28148966) »
  • Park MJ  et al. Lipotoxicity-Induced PRMT1 Exacerbates Mesangial Cell Apoptosis via Endoplasmic Reticulum Stress. Int J Mol Sci 18:N/A (2017). Read more (PubMed: 28671608) »

See all 47 Publications for this product

Customer reviews and Q&As

Thank you for getting back to me. I am sorry to hear that your customer is still dissatisfied with the quality of this antibody. Please do let me know how you wish to proceed with this enquiry. I could offer you either a free of charge replacement vial...

Read More
Application
Western blot
Sample
Pig Cell lysate - whole cell (vascular endothelial cell)
Gel Running Conditions
Non-reduced Denaturing (6% gel)
Loading amount
30 µg
Specification
vascular endothelial cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted May 05 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (N87 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel)
Loading amount
800000 cells
Specification
N87 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Aug 03 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Panc1 pancreatic cancer cell line)
Permeabilization
Yes - 0.25% TritonX-100 in PBS 10 minutes
Specification
Panc1 pancreatic cancer cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 19 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (retina)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
40 µg
Specification
retina
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jul 20 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mesenteric artery)
Permeabilization
No
Specification
mesenteric artery
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Jul 06 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Mouse Cell (retina)
Specification
retina
Permeabilization
Yes - Triton X-100
Fixative
Paraformaldehyde
Username

Dr. Minzhong Yu

Verified customer

Submitted Jan 07 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12% gel, TGS buffer)
Sample
Mouse Tissue lysate - whole (Heart left ventricule)
Specification
Heart left ventricule
Treatment
Tunicamycin for 48h
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Dec 19 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Non-reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (prostate cancer cell 22Rv1)
Specification
prostate cancer cell 22Rv1
Treatment
300nM Thapsigargin for 6 and 12 hours
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 20 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (retina)
Loading amount
30 µg
Specification
retina
Treatment
25mM high glucose
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Mr. Dongil Kim

Verified customer

Submitted Apr 11 2013

1-10 of 20 Abreviews or Q&A

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