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PDP - Template Name: Antibody Sampler Kit
PDP - Template ID: *******4a3ef3a
  1. Products /
  2. Primary Antibodies /
  3. Exosomal Marker Antibody Sampler Kit

Exosomal Marker Antibody Sampler Kit #74220

    Product Information

    Kit Usage Information

    Protocols

    Product Description

    The Exosomal Marker Antibody Sampler Kit provides an economical means to evaluate the presence of exosomal markers. The kit includes enough primary antibody to perform two western blot experiments for each target.

    Specificity / Sensitivity

    All antibodies provided in the kit detect endogenous levels of the respective target protein. Additionally, the Annexin V Antibody is not predicted to cross-react with other annexin family members and the GM130 antibody may cross-react with a protein of unknown origin at 30 kDa.

    Source / Purification

    Monoclonal antibodies are produced by immunizing animals with full-length recombinant human Alix protein, a synthetic peptide corresponding to residues surrounding Pro410 of human CD54/ICAM-1 protein, a synthetic peptide corresponding to residues surrounding Val178 of human CD9 protein, residues surrounding Thr195 of human GM130 protein, residues near the amino terminus of human EpCAM protein, and residues surrounding Ile368 of human flotillin-1 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human annexin V protein and residues surrounding Asp69 of human HSP70. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

    Background

    Exosomes are small membrane-bound vesicles that in recent years have emerged as important molecules for inter-cellular communication. Exosomes are produced during both normal and pathophysiological conditions, and cancer cells have been shown to secrete exosomes in greater amounts than normal cells (reviewed in 1). The exosomal markers contained in this kit are Alix, Annexin V, ICAM-1, CD9, GM130, EpCAM, flotillin, and HSP70.
    Alix, a cytosolic scaffold protein, regulates many cellular processes including endocytic membrane trafficking, cell adhesion through interactions with ESCRT (endosomal sorting complex required for transport) proteins, endophilins, and CIN85 (Cbl-Interacting protein of 85 kDa) (2, 3).
    Annexin V is a ~30 kDa protein that binds to phospho-lipids in a calcium-dependent manner (4). All annexins contain a putative PKC binding site, but only annexin V has been identified as an inhibitor of this pathway (5).
    Intracellular cell adhesion molecule-1 (CD54 or ICAM-1) is a cell surface glycoprotein that belongs to the immunoglobulin superfamily (IgSF) of adhesion molecules. CD54 is expressed at low levels in diverse cell types, and is induced by cytokines (TNF-alpha, interleukin-1) and bacterial lipopolysaccharides (6). Apical localization on endothelial cells (or basolateral localization on epithelial cells) is a prerequisite for leukocyte trafficking through the endothelial (or epithelial) barrier (6).
    The CD9 antigen belongs to the tetraspanin family of cell surface glycoproteins. Tetraspanins interact with a variety of cell surface proteins and intracellular signaling molecules in specialized tetraspanin-enriched microdomains (TEMs), where they mediate a range of processes including adhesion, motility, membrane organization, and signal transduction (7). Additional research identified CD9 as an abudant component of exosomes, and may play a role in the fusion of these secreted membrane vesicles with recipient cells (8).
    GM130 is required for membrane fusion events that mediate ribbon formation during Golgi assembly (9). The Golgi apparatus functions in the modification, organization, and transport of proteins and membrane targeted to other parts of the cell, such as the plasma membrane, lysosomes, and endosomes. This regulated transport is important for appropriate protein localization, secretion, and signal transduction (reviewed in 10).
    Epithelial cell adhesion and activation molecule (EpCAM/CD326) is a transmembrane glycoprotein that mediates calcium-independent, hemophilic adhesions on the basolateral surface of most epithelial cells (11). One of the first tumor-associated antigens discovered, EpCAM has long been a marker of epithelial and tumor tissue. Research studies have shown that EpCAM is highly expressed in cancer cells and can be used as a biomarker for the detectionof tumor-derived exposomes (reviewed in 1, 12, 13).
    Flotillins belong to a famiy of lipid raft-associated integral membrane proteins that are ubiquitously expressed and located to lipid rafts on the cell plasma membrane where they support signal transduction and regulate lipid raft motility and localization (14-17). In addition to its colocalization with lipid rafts on the plasma membrane, flotillin-1 also has been found at compartments of the endocytic and autophagosomal pathways, such as recycling/ late endosomes, the Golgi complex, as well as the nucleus (18, 19).
    HSP70 is a molecular chaperone expressed constituitively under normal conditions to maintain protein homeostatis and is induced upon environmental stress (20). HSP70 is able to interact with unfolded proteins to prevent irreversible aggregation and catalyze the refolding of their substrates in an ATP and co-chaperone dependent manner (21). An immune response is elicited upon excretion of heat shock proteins from tumor exosomes (reviewed in 1).

    1. Raposo, G. and Stoorvogel, W. (2013) J Cell Biol 200, 373-83.
    2. Katoh, K. et al. (2003) J Biol Chem 278, 39104-13.
    3. Sadoul, R. (2006) Biol Cell 98, 69-77.
    4. Huber, R. et al. (1990) EMBO J 9, 3867-74.
    5. Cardó-Vila, M. et al. (2003) Mol Cell 11, 1151-62.
    6. Hopkins, A.M. et al. (2004) Adv Drug Deliv Rev 56, 763-78.
    7. Hemler, M.E. (2005) Nat Rev Mol Cell Biol 6, 801-11.
    8. Théry, C. et al. (1999) J Cell Biol 147, 599-610.
    9. Puthenveedu, M.A. et al. (2006) Nat Cell Biol 8, 238-48.
    10. Barr, F.A. and Short, B. (2003) Curr Opin Cell Biol 15, 405-13.
    11. Went, P.T. et al. (2004) Hum Pathol 35, 122-8.
    12. Baeuerle, P.A. and Gires, O. (2007) Br J Cancer 96, 417-23.
    13. Armstrong, A. and Eck, S.L. Cancer Biol Ther 2, 320-6.
    14. Langhorst, M.F. et al. (2005) Cell Mol Life Sci 62, 2228-40.
    15. Stuermer, C.A. and Plattner, H. (2005) Biochem Soc Symp , 109-18.
    16. Fernow, I. et al. (2007) Eur J Cell Biol 86, 345-52.
    17. Neumann-Giesen, C. et al. (2007) J Cell Sci 120, 395-406.
    18. Liu, J. et al. (2005) J Biol Chem 280, 16125-34.
    19. Santamaría, A. et al. (2005) Mol Cell Biol 25, 1900-11.
    20. Nollen, E.A. and Morimoto, R.I. (2002) J Cell Sci 115, 2809-16.
    21. Young, J.C. et al. (2003) Trends Biochem Sci 28, 541-7.

    Database Links

    UniProt ID: P05362 , Q08379 , P16422 , O75955 , P21926 , P08758 , P0DMV8 , Q8WUM4


    Entrez-Gene Id: 3383 , 2801 , 4072 , 10211 , 928 , 308 , 3303 , 10015


    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    For Research Use Only. Not For Use In Diagnostic Procedures.
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