Invitrogen

Phospho-IRE1 alpha (Ser724) Polyclonal Antibody

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Phospho-IRE1 alpha (Ser724) Antibody in Western Blot (WB) — Western blot analysis of IRE1 alpha (Ser724) in Min6 cells treated with different concentrations of glucose for 3 hours prior to lysates preparation using anti-IRE1 polyclonal antibody (Product # PA1-16927).

Phospho-IRE1 alpha (Ser724) Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of IRE1 alpha (Ser724) in immersion-fixed paraffin-embedded sections of human spleen using a polyclonal anti-IRE1 S724 antibody (Product # PA1-16927) at 1:300 for 1 hour at room temperature, followed by incubation with an anti-Rabbit IgG HRP polymer antibody. Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the perinuclear cytoplasm in splenocytes.

Phospho-IRE1 alpha (Ser724) Antibody — Fig. 1 PERK-mediated secretion of proteins under hypoxia. a Total PERK, eIF2alpha and P-eIF2alpha expression in HEK293, LN308 and LN229 cell lines under normoxia or hypoxia. b Active form of ATF6alpha (50 kDa band) in LN308, under hypoxia for 24, 48 and 72 h. EEF2 was used as a loading control. c Relative mRNA levels of XBP1s as determined by qRT-PCR in LN308 and LN229 cell lines under hypoxia (48 h). NDRG1 was taken as a positive control for hypoxia induction. EEF2 was used as housekeeping gene. Data are normalized to the respective normoxic conditions and are represented as the mean of three independent experiments +- SEM ( T test: ** p value < 0.01). N normoxia, H hypoxia. d Total XBP1s mRNA transcripts in HEK293 and LN308 cells treated with hypoxia for 24, 48, 72 h. beta-actin was used as a housekeeping gene. e Total P-IRE1alpha species immunoprecipitated using P-IRE1alpha antibody from HEK293 and LN308 cells treated with hypoxia for 48 h. f Volcano plot representing the regulated secretory proteins from LN308 glioblastoma cells under hypoxic conditions for 72 h without (left) and with PERK inhibitor (GSK2606414; right). The data are represented as the mean of three independent replicates. The significant p value cut-off was set at 0.05.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1000

View 35 publications 35 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:10-1:500

Immunohistochemistry (Frozen) (IHC (F))

1:10-1:500

Flow Cytometry (Flow)

View 1 publication 1 publication

ELISA (ELISA)

1:100-1:2000

Immunoprecipitation (IP)

1:10-1:500

View 1 publication 1 publication

ChIP assay (ChIP)

Assay Dependent

Immunocytochemistry (ICC/IF)

1:10-1:500

View 1 publication 1 publication

Miscellaneous PubMed (Misc)

View 3 publications 3 publications

Product Specifications

Species Reactivity

Fruit fly, Goat, Human, Mouse, Non-human primate, Pig, Rabbit, Rat

Published species

Human, Mouse, Rat

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide surrounding the phosphorylated serine 724 of the human IRE1 alpha protein.[Swiss-Prot #O75460]

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS

Contains

0.025% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2262241

Product Specific Information

Recommended positive controls: Human pancreas whole tissue Lysate (adult whole tumor), Human epididymis whole tissue lysate (adult whole normal)

Target Information

IRE1 alpha is the endoplasmic reticulum to nucleus signalling 1 protein, a human homologue of the yeast Ire1 gene product. IRE1 alpha protein possesses intrinsic kinase activity and an endoribonuclease activity and it is important in altering gene expression as a response to endoplasmic reticulum-based stress signals. Specifically, IRE1 acts as the sensor of unfolded proteins in the ER that initiates transmittance of the unfolded protein signal from the ER to the nucleus by splicing XBP1 mRNA converting it into a potent unfolded-protein response transcriptional activator. Null mutants are viable, but IRE1 is essential for viability under stress conditions that cause unfolded proteins to accumulate in the ER. Depletion of IRE1p through the expression of a dominant negative form of IRE1p has no effect on transfected cells, but cell death via apoptosis occurs under stress conditions that cause unfolded proteins to accumulate in the ER.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: endoplasmic reticulum (ER) to nucleus signalling 1; endoplasmic reticulum to nucleus signalling 1; Endoplasmic reticulum-to-nucleus signaling 1; Endoribonuclease; ER to nucleus signalling 1; ERN1; FLJ30999; hIRE1p; inositol-requiring 1; inositol-requiring 1 alpha; inositol-requiring enzyme 1; Inositol-requiring protein 1; Ire1-alpha; IRE1a; IRE1a antibody; IRE1p antibody; MGC163277; MGC163279; protein kinase/endoribonuclease; Serine/threonine-protein kinase; Serine/threonine-protein kinase/endoribonuclease IRE1

Gene Aliases: 9030414B18Rik; AI225830; C85377; ERN1; hIRE1p; IRE1; IRE1a; Ire1alpha; IRE1P; RGD1559716

UniProt ID: (Human) O75460, (Mouse) Q9EQY0

Entrez Gene ID: (Rabbit) 100357346, (Goat) 102173038, (Human) 2081, (Pig) 100510978, (Mouse) 78943, (Rat) 498013

Molecular Function: kinase protein kinase transferase