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

Description

• General description: We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
• 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 TestingEvaluated 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 applicationsWestern 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
• 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.