Revision 2

#80610Store at -20C

1 Kit

(7 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
CD9 (D8O1A) Rabbit mAb 13174 20 µl 22, 24, 35 kDa Rabbit IgG
CD81 (D3N2D) Rabbit mAb 56039 20 µl 22 kDa Rabbit IgG
CD63 (E1W3T) Rabbit mAb 52090 20 µl 25-60 kDa Rabbit IgG
TSG101 (E6V1X) Rabbit mAb 72312 20 µl 50 kDa Rabbit IgG
Alix (E6P9B) Rabbit mAb 92880 20 µl 90-100 kDa Rabbit IgG
Flotillin-1 (D2V7J) XP® Rabbit mAb 18634 20 µl 49 kDa Rabbit IgG
Syntenin-1/MDA9 (E2I9L) Rabbit mAb 27964 20 µl 30 kDa Rabbit IgG
HSP70 Antibody 4872 20 µl 72, 73 kDa Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Human Reactive Exosome Marker Antibody Sampler Kit provides an economical means of analyzing proteins that can be present on exosomes. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Exosomes are small (30-150 nm) membrane-bound vesicles that are secreted by various cell types under normal and pathological conditions (1,2). They originate from intracellular multivesicular endosomes upon fusion with the plasma membrane. Exosomes have emerged as an important mechanism of intercellular communication facilitating the transfer of membrane and cytosolic proteins, lipids, and RNA.

A variety of methods have been described to isolate exosomes and understand their composition (3-7). Heterogeneity in exosome composition can be attributed to the cells of origin as well as the isolation methods. However, there are protein markers that appear with high frequency. Tetraspanins are a family of cell surface glycoproteins with four transmembrane domains often found in exosomes (8). Tetraspanins CD9, CD81, and CD63 appear in exosomes and have been the target of immune-affinity approaches of exosome isolation. Flotillin-1 is a lipid raft-associated integral membrane protein that is incorporated into exosomes (9). Exosomes also contain proteins involved in endosomal membrane trafficking, collectively known as the ESCRT (endosomal sorting complex required for transport) pathway. Alix regulates cellular processes, such as endocytic membrane trafficking and cell adhesion through interactions with ESCRT proteins including endophilins, and CIN85 (Cbl-interacting protein of 85 kDa), and plays a role in exosome biogenesis (10-12). Syntenin-1 (MDA9, SDCBP) is a member of the PDZ family of proteins that functions as a scaffold adaptor protein regulating numerous signal transduction pathways (13). Syntenin-1 interacts with Alix to regulate exosome biogenesis (12). Tumor susceptibility gene 101 (TSG101) is a fundamental component of the ESCRT complex I involved in regulating the trafficking of proteins throughout the endosomal compartment (14). TSG101 is involved in regulating diverse biological processes, such as cell proliferation, viral budding and release, and exosome biosynthesis (15,16). The heat shock protein HSP70 is a molecular chaperone involved in protein folding that can be induced upon environmental stress (17). HSP70 may also be secreted through exosomes (18).

  1. Raposo, G. and Stoorvogel, W. (2013) J Cell Biol 200, 373-83.
  2. van Niel, G. et al. (2018) Nat Rev Mol Cell Biol 19, 213-228.
  3. Jeppesen, D.K. et al. (2019) Cell 177, 428-445.e18.
  4. Kowal, J. et al. (2016) Proc Natl Acad Sci U S A 113, E968-77.
  5. Sidhom, K. et al. (2020) Int J Mol Sci 21, 6466. doi: 10.3390/ijms21186466.
  6. Patel, G.K. et al. (2019) Sci Rep 9, 5335.
  7. Tauro, B.J. et al. (2012) Methods 56, 293-304.
  8. Hemler, M.E. (2005) Nat Rev Mol Cell Biol 6, 801-11.
  9. de Gassart, A. et al. (2003) Blood 102, 4336-44.
  10. Katoh, K. et al. (2003) J Biol Chem 278, 39104-13.
  11. Chatellard-Causse, C. et al. (2002) J Biol Chem 277, 29108-15.
  12. Baietti, M.F. et al. (2012) Nat Cell Biol 14, 677-85.
  13. Pradhan, A.K. et al. (2020) Cancer Metastasis Rev 39, 769-781.
  14. Katzmann, D.J. et al. (2001) Cell 106, 145-55.
  15. Garrus, J.E. et al. (2001) Cell 107, 55-65.
  16. Zhong, Q. et al. (1998) Cancer Res 58, 2699-702.
  17. Nollen, E.A. and Morimoto, R.I. (2002) J Cell Sci 115, 2809-16.
  18. Zhan, R. et al. (2009) Biochem Biophys Res Commun 387, 229-33.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
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