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Wählen Sie konfigurierbare Panels & Premixed-Kits - ODER - Kits für die zelluläre Signaltransduktion & MAPmates™
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Konfigurierbare Panels & Premixed-Kits
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Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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Gewähltes Kit
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96-Well Plate
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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Programmed 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
Format
Purified
Presentation
Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Anti-Alix Antibody, clone 1A3 is an antibody against Xp95 for use in Western Blotting, Immunoprecipitation, Immunocytochemistry.
Key Applications
Western Blotting
Immunoprecipitation
Immunocytochemistry
Application Notes
Immunoprecipitation 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
Immunogen
GST-tagged recombinant full-length human Alix.
Epitope
Patch 2 of the Bro1 domain.
Clone
1A3
Concentration
Please refer to lot specific datasheet.
Host
Mouse
Specificity
Clone 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).
Isotype
IgG1κ
Species Reactivity
Human
Xenopus
Species Reactivity Note
Human, Xenopus. Predicted to react with Mouse based on 100% sequence homology.
~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 Assurance
Evaluated 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.