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96128
Mouse Reactive Exosome Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Mouse Reactive Exosome Marker Antibody Sampler Kit #96128

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Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using CD81 (D5O2Q) Rabbit mAb (Mouse Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 2
Western blot analysis of extracts from various cell lines using Flotillin-1 (D2V7J) XP® Rabbit mAb.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 3
Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using HSP70 Antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 4
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 5
Western blot analysis of extracts from various cell lines using TSG101 (E6V1X) Rabbit mAb.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 6
Western blot analysis of extracts from HCT 116 and HCT 116 Alix knockout (-/-) cells using Alix (E6P9B) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 7
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse CD9 protein (mCD9-Myc/DDK; +), using CD9 (E8L5J) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 8
Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse CD81 (mCD81-Myc/DDK, +), using CD81 (D5O2Q) Rabbit mAb (Mouse Specific).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 9
Immunoprecipitation of flotillin-1 protein from BT-20 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Flotillin-1 (D2V7J) XP® Rabbit mAb. Western blot analysis was performed using Flotillin-1 (D2V7J) XP® Rabbit mAb.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 10
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using HSP70 Antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 11
Western blot analysis of extracts from 293T cells, transfected with control siRNA (-) or TSG101 siRNA (+), using TSG101 (E6V1X) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 12
Western blot analysis of extracts from various cell lines using Alix (E6P9B) Rabbit mAb.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 13
Western blot analysis of extracts from various cell lines using CD9 (E8L5J) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Absence of signal in BA/F3 cells is predicted from RNAseq data and confirms the specificity of the antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 14
Immunoprecipitation of CD81 from extracts of C2C12 cells. Lane 1 is 10% input, Lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is CD81 (D5O2Q) Rabbit mAb (Mouse Specific). Western blot analysis was performed using CD81 (D5O2Q) Rabbit mAb (Mouse Specific).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using HSP70 Antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 16
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human TSG101 (hTSG101-Myc/DDK; +), using TSG101 (E6V1X) Rabbit mAb (upper), DYKDDDDK Tag (D6W5B) Rabbit mAb #14793 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 17
Immunoprecipitation of Alix from K-562 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control, and lane 3 is Alix (E6P9B) Rabbit mAb. Western blot was performed using Alix (E6P9B) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was using as a secondary antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 18
Confocal immunofluorescent analysis of RAW 264.7 cells (left, positive) or BA/F3 cells (right, negative) using CD9 (E8L5J) Rabbit mAb (green) and DAPI #4083 (blue).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 19
Confocal immunofluorescent analysis of C2C12 (positive, left) and BaF3 (negative, right) cells using CD81 (D5O2Q) Rabbit mAb (Mouse Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Flotillin-1 (D2V7J) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded human lung carcinoma showing cytoplasmic localization using HSP70 Antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 22
Confocal immunofluorescent analysis of HCT 116 cells (left, positive) and HCT 116 Alix knockout (-/-) cells (right, negative) using Alix (E6P9B) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 23
Flow cytometric analysis of live BA/F3 cells (blue, negative) and JAWSII cells (green, positive) using CD9 (E8L5J) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using HSP70 Antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 25
Flow cytometric analysis of fixed and permeabilized BA/F3 cells (blue, negative) and JAWSII cells (green, positive) using CD9 (E8L5J) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 26
Confocal immunofluorescent analysis of BT-20 cells using Flotillin-1 (D2V7J) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using HSP70 Antibody.
Mouse Reactive Exosome Marker Antibody Sampler Kit: Image 28
Flow cytometric analysis of live EL4 cells (blue, negative) and A20 cells (green, positive) using CD81 (D5O2Q) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 96128
Cat. # Size Qty. Price
96128T
1 Kit  (6 x 20 microliters)
$ 518

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CD9 (E8L5J) Rabbit mAb 98327 20 µl
  • WB
  • IF
  • F
M R 22-27 Rabbit IgG
CD81 (D5O2Q) Rabbit mAb (Mouse Specific) 10037 20 µl
  • WB
  • IP
  • IF
  • F
M 22 Rabbit IgG
TSG101 (E6V1X) Rabbit mAb 72312 20 µl
  • WB
H M R 50 Rabbit IgG
Alix (E6P9B) Rabbit mAb 92880 20 µl
  • WB
  • IP
  • IF
H M R 90-100 Rabbit IgG
Flotillin-1 (D2V7J) XP® Rabbit mAb 18634 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 49 Rabbit IgG
HSP70 Antibody 4872 20 µl
  • WB
  • IHC
H M R Mk B 72, 73 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The Mouse 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.

Specificity / Sensitivity

Each antibody in the Mouse Reactive Exosome Marker Antibody Sampler Kit detects endogenous levels of its target protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu119 of mouse CD9, Pro184 of mouse CD81, Pro346 of human Alix, Ile368 of human flotillin-1, and residues near the amino terminus of human TSG101. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to human HSP70. Antibodies are purified by protein A and peptide affinity chromatography.

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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