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MABS1277 Anti-Alix/Xp95 Antibody, clone 1A3

Anti-Alix Antibody, clone 1A3 is an antibody against Xp95 for use in Western Blotting, Immunoprecipitation, Immunocytochemistry.

Anti-Alix/Xp95 Antibody, clone 1A3 MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Signaling Research Sub-Category: Vesicular Trafficking Gene Symbol: PDCD6IP, AIP1, ALIX, KIAA1375 UniProt Number: Q8WUM4
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MABS1277
100 μg  
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      Overview

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      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, XnWB, IP, ICCMPurifiedMonoclonal Antibody
      Description
      Catalogue NumberMABS1277
      DescriptionAnti-Alix/Xp95 Antibody, clone 1A3
      Alternate Names
      • Programmed cell death 6-interacting protein
      • ALG-2-interacting protein 1
      • ALG-2-interacting protein X
      • Apoptosis-linked gene-2-interacting protein X
      • Hp95
      • Human ortholog of Xenopus protein of 95 kDa
      • Human ortholog of Xp95
      • PDCD6-interacting protein
      Background InformationProgrammed cell death 6-interacting protein (UniProt Q8WUM4; also known as ALG-2-interacting protein 1, ALG-2-interacting protein X, Apoptosis-linked gene-2-interacting protein X, Hp95, Human ortholog of Xenopus protein of 95 kDa, Human ortholog of Xp95, PDCD6-interacting protein) is encoded by the PDCD6IP (also known as AIP1, ALIX, KIAA1375) gene (Gene ID 10015) in human. Alix, the mammalian orthologue of xenopus Xp95 and yeast BRO1, is an abundant adaptor protein involved in diverse cellular processes, including actin-based cytoskeleton assembly, integrin-mediated cell adhesion, and ESCRT- (endosomal sorting complex required for transport-) dependent membrane trafficking. Sorting of endocytosed membrane proteins by multivesicular bodies (MVBs) requires coordinated cargo recognition and assembly of membrane-sculpturing machinery that buds cargo-loaded membrane vesicles into the lumen of endosomes. MVB-mediated sorting is driven by five distinct ESCRT complexes (ESCRT-0, -I, -II and -III, and the Vps4 complex) and associated proteins. Alix and charged MVB protein 4 (CHMP4), the mammalian orthologue of yeast Snf7, are binding partners involved in a variety of ESCRT-III-mediated membrane-remodelling processes in mammalian cells, including retroviral budding, cytokinetic abscission, biogenesis of exosomes, plasma membrane repair, and ubiquitin-independent MVB sorting of protease-activated receptor 1 (PAR1). In the absence of binding partners, an intramolecular interaction between the Patch 2 region in the Bro1 domain (a.a. 3-392) and the TSG101-docking site (a.a. 717-720) in the Pro-rich domain (a.a. 717-860) locks Alix in a conformation inaccessible for interaction with CHMP4 and retroviral Gag proteins.
      References
      Product Information
      FormatPurified
      PresentationPurified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationAnti-Alix Antibody, clone 1A3 is an antibody against Xp95 for use in Western Blotting, Immunoprecipitation, Immunocytochemistry.
      Key Applications
      • Western Blotting
      • Immunoprecipitation
      • Immunocytochemistry
      Application NotesImmunoprecipitation Analysis: A representative lot immunoprecipiated native Alix (1-709) GST fusion with C-terminal proline-rich domain (PRD; a.a. 710-868) deletion, but not native full-length Alix (1-868) GST fusion or native Alix (1-746) GST fusion with only partial PRD deletion (Zhou, X., et al. (2010). Biochem. J. 432(3):525-534).
      Immunoprecipitation Analysis: A representative lot immunoprecipiated GST-Bro1 domain fragment, but not GST-full-length Alix protein under native conditions. Clone 1A3 immunoprecipitated full-length Alix from HEK293 lysates prepared with RIPA buffer, 1% NP40, or 0.1% SDS, but not from lysates prepared with TBS, 0.5% DOC, or 0.1% Triton X-100 (Zhou, X., et al. (2009). Biochem. J. 418(2):277-284).
      Western Blotting Analysis: A representative lot detected wild-type human Alix and Xenopus Xp95 Bro1 domain GST fusions, but not the corresponding GST fusions with Y319F mutation in the Patch2 region (Zhou, X., et al. (2009). Biochem. J. 418(2):277-284).
      Western Blotting Analysis: A representative lot detected recombinant xenopus Xp95 and human Alix GST fusion proteins, as well as endogenous Xp95 in extracts of both mature and immature Xenopus oocytes (Zhou, X., et al. (2009). Biochem. J. 418(2):277-284).
      Western Blotting Analysis: A representative lot detected recombinant human Alix fragment a.a. 168-436, but not a.a. 436-709 (Pan, S., et al. (2008). EMBO J. 27(15):2077-2090).
      Immunocytochemistry Analysis: A representative lot immunostained extracellular clumps and fibers by fluorescent immunocytochemistry staining of 4% paraformaldehyde-fixed, 0.5% Triton X-100-permeabilized WI-30 human diploid fetal lung fibroblasts. Alix immunostaining overlapped with that of anti-fibronectin, siRNA-mediated Alix knockdown eliminated the staining by clone 1A3 (Pan, S., et al. (2008). EMBO J. 27(15):2077-2090).
      Note: In the native conformation of a full-length Alix protein, the Bro1 domain Patch 2 region is not exposed due to intramolecular interaction with the C-terminal proline-rich domain (PRD). RIPA buffer, 1% NP40, or 0.1% SDS (but not 0.5% DOC, or 0.1% Triton X-100) have been shownn to effectly alter Alix conformation and expose the Patch 2 region for antibody binding and protein interaction in immunoprecipitation and affinity interaction applications (Zhou, X., et al. (2009). Biochem. J. 418(2):277-284).
      Biological Information
      ImmunogenGST-tagged recombinant full-length human Alix.
      EpitopePatch 2 of the Bro1 domain.
      Clone1A3
      ConcentrationPlease refer to lot specific datasheet.
      HostMouse
      SpecificityClone 1A3 recognizes an epitope conserved among human Alix and xenopus Xp95, localized within the Bro1 domain C-terminal end Patch 2 region (Zhou, X., et al. (2009). Biochem. J. 418(2):277-284). Clone 1A3 immunostained WI-30 cells before, but not after, siRNA-mediated Alix knockdown (Pan, S., et al. (2008). EMBO J. 27(15):2077-2090). Epitope region is present in human Alix spliced isoform 1 and 2 (equivalent to mouse isoform 1 and 3, respectively), but absent in isoform 3 (equivalent to mouse isoform 2) reported by UniProt (Q8WUM4 for human and Q9WU78 for mouse).
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Xenopus
      Species Reactivity NoteHuman, Xenopus. Predicted to react with Mouse based on 100% sequence homology.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • PDCD6IP
      • AIP1
      • ALIX
      • KIAA1375
      Purification MethodProtein G Purified
      UniProt Number
      Molecular Weight~96 kDa observed. 96.02 kDa (human & mouse isoform 1), 96.77 kDa (human isoform 2 & mouse isoform 3) calculated. Uncharacterized band(s) may appear in some lysates.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting in Daudi cell lysate.

      Western Blotting Analysis: 0.5 µg/mL of a representative lot detected Alix in 10 µg of Daudi cell lysate.
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
      Packaging Information
      Material Size100 μg
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      MABS1277 04055977350104