ZRB1072

Sigma-Aldrich

Anti-phospho-IRE-1α (Ser724) Antibody, clone 1M2 ZooMAb® Rabbit Monoclonal

newenhanced validation

recombinant, expressed in HEK 293 cells

Quality Level

biological source

rabbit

recombinant

expressed in HEK 293 cells

antibody form

purified antibody

antibody product type

primary antibodies

clone

1M2, recombinant monoclonal

description

recombinant, expressed in HEK 293 cells

product line

ZooMAb® learn more

form

lyophilized

mol wt

calculated mol wt 109.74 kDa
observed mol wt ~110 kDa

species reactivity

mouse, human

species reactivity (predicted by homology)

rat

packaging

antibody small pack of 25 μL

enhanced validation

recombinant expression
Learn more about Antibody Enhanced Validation

application(s)

affinity binding assay: suitable
immunocytochemistry: suitable
peptide synthesis: suitable
western blot: suitable

isotype

IgG

epitope sequence

C-terminal half

conjugate

unconjugated

Protein ID accession no.

UniProt accession no.

shipped in

ambient

storage temp.

2-8°C

General description

ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.

Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.

Learn more about ZooMAb here.

Specificity

Clone IM2 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Serine/threonine-protein kinase/endoribonuclease IRE1 (IRE-1a) phosphorylated on serine 724.

Immunogen

KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphoserine 724 from the C-terminal, cytoplasmic domain of human Serine/threonine-protein kinase/endoribonuclease IRE1 (IRE-1a).

Application

Quality Control Testing

Evaluated by Western Blotting in lysate from HeLa cells treated with Calyculin A.

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected phospho-IRE- 1a (Ser724) lysate from NIH3T3 cells treated with Calyculin A (50 nM; 30 min.).

Tested applications

Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected phospho-IRE-1a (Ser724) in untreated NIH3T3 and NIH3T3 treated with Calyculin A.

Affinity Binding Assay: A representative lot of this antibody bound IRE-1a (phospho Ser724) peptide with a KD of 9.5 x 10-8 in an affinity binding assay.

Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected phospho-IRE-1a (Ser724) in C2C12 cells..

Peptide Inhibition Assay: Target band detection in lysate from NIH3T3 cells treated with 50 nM Calyculin A was prevented by preblocking of a representative lot of phospho-IRE-1a (Ser724) with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Anti-phospho-IRE-1a (Ser724), clone 1M2 ZooMAb®, Cat. No. ZRB1072, is a Rabbit monoclonal antibody that targets phospho-IRE1a (Ser724) and is tested for use in Affinity Binding Assay, Immunocytochemistry, Peptide Inhibition Assay, and Western Blotting.

Target description

Serine/threonine-protein kinase/endoribonuclease IRE1 (UniProt: O75460; also known as Endoplasmic reticulum-to-nucleus signaling 1, Inositol-requiring protein 1, hIRE1p, Ire1-alpha, IRE 1a ) is encoded by the ERN1 (also known a IRE1) gene (Gene ID: 2081) in human. IRE1a is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-18), which is subsequently cleaved off to generate the mature form that contains a lumenal domain (aa 19-443), a transmembrane domain (aa 444-464), and a long cytoplasmic domain (aa 465-977). It serves as a serine/threonine-protein kinase and endoribonuclease that acts as a key sensor for the endoplasmic reticulum unfolded protein response (UPR). IRE1 activity can induce both protective UPR and apoptotic downstream signaling events. In unstressed cells, the endoplasmic reticulum luminal domain is maintained in its inactive monomeric state by binding to the endoplasmic reticulum chaperone HSPA5/BiP. Accumulation of misfolded proteins in the endoplasmic reticulum causes release of HSPA5/BiP, which allows the luminal domain to homodimerize and promote autophosphorylation of the kinase domain and subsequent activation of its endoribonuclease activity. Its endoribonuclease activity is shown to be specific for XBP1 mRNA and excises 26 nucleotides from XBP1 mRNA. The resulting spliced transcript of XBP1 encodes a transcriptional activator protein that up-regulates expression of UPR target genes. IRE1a is shown to induce transient ER stress signaling and confer a protective effect against pressure overload-induced pathological remodeling in the murine heart. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Steiger, D., et al (2018). J. Biol. Chem. 293(25); 9652-9661).

Physical form

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.

Reconstitution

300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Legal Information

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis
Certificate of Origin

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