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High Quality anti-Human Tetraspanin Monoclonal Antibodies

For characterizing and isolating classical exosomes using new “ultraclean” methods.

This product is an antibody that specifically recognizes CD9, CD63, and CD81, known as exosome markers, and can isolate exosomes from serum and culture supernatants by immunoprecipitation.

Features

  • ●High Specificity for tetraspanin proteins, CD9, CD63 and CD81
  • ●Sample Type (Human samples)
    • CD9 : serum, plasma, Cell culture supernatants and urine
    • CD63 : serum, plasma, Cell culture supernatants and urine
    • CD81 : serum, plasma and Cell culture supernatants
  • ●CD9 Antibody was tedted with bovine milk exosomes and CD81 antibody was tested with bovine milk exosomes and FBS.
  • ●Analyisis of exosome surface antigens, exosomal miRNAs and proteins included in exosomes

Data

IP-WB

1. IP-WB of CD9 in serum Exosome with anti CD9 (12A12) antibody
1. CD9 antibody (12A12) Serum A
2. control antibody Serum A
3. CD9 antibody (12A12) Serum B
4. control antibody Serum B
5. ultracentrifugation Serum A
6. ultracentrifugation Serum B
2. IP-WB of CD63 in serum Exosome with anti CD63 (8A12) antibody
1. CD63 antibody (8A12) Serum A
2. control antibody Serum A
3. CD63 antibody (8A12) Serum B
4. control antibody Serum B
5. ultracentrifugation Serum A
6. ultracentrifugation Serum B
3. IP-WB of CD81 in serum Exosome with anti CD81 (12C4) antibody
1. CD81 antibody (12C4) Serum A
2. control antibody Serum A
3. CD81 antibody (12C4) Serum B
4. control antibody Serum B
5. ultracentrifugation Serum A
6. ultracentrifugation Serum B

Immunofluorescent Cell Staining (ICC / IF)

Localization of CD63 in prostate cancer cell line PC-3M and colon cancer cell line HCT116 using anti CD63 (8A12) antibody
  • A: prostate cancer cell line, PC-3M
  • B: colon cancer cell line, HCT116

Secondary antibody is Alexa Fluor(R) 488-labeled anti mouse secondary antibody.

Images courtesy of :
National Cancer Center Research Institute of Molecular and Cellular Therapy
Tokyo Medical University

Immunoelectron Microscopy

Immunoelectron microscopic detection of human CD9 and CD63 protein on exosome surfaces
  • (A) Anti-human CD9 antibody (clone 12A12) detected CD9 on the surface of exosomes from human breast cancer cells.
  • (B) Anti-human CD63 antibody (clone 8A12) also detected CD63 on the surface of exosomes from human breast cancer cells.
Images courtesy of :
National Cancer Center Research Institute of Molecular and Cellular Therapy

Cryo-electron microscopy

Cryo-electron microscopic detection of exosomes bound by anti-human CD63 antibody (clone 8A12).

Exosomes and anti-human CD63 antibody (clone 8A12) were mixed and imaged with cryo-electron microscopy (Titan Krios). Shown is a cryo-electron micrograph of an exosome (A), a three-dimensional reconstruction of an exosome (green) with a single bound antibody (cyan) (B) and a rotating 3D tomographic reconstruction of an exosome and a bound antibody (C).

Images courtesy of :
Okinawa Protein Tomography Ltd.

Specification

TargetCloneImmunogenReactiveNon-reactiveApplicationIsotype
CD9 12A12 Amino acids 113-195: the large second extracellular loop between the 3rd and 4th transmembrane domains Human Mouse, Rat IP, WB, FC, ELISA, IF, IHC, IEM IgG2b
CD63 8A12 Amino acids 104-202: the large second extracellular loop between the 3rd and 4th transmembrane domains Human Mouse, Rat IP, WB, FC, ELISA, IF, IHC, IEM IgG2a
CD81 12C4 Amino acids 36-54: the small first extracellular loop between the 3rd and 4th transmembrane domains Human, Bovine Mouse, Rat IP, WB, FC, ELISA, IF, IHC, IEM IgG2a

＊CD9 antibody and CD63 antibody were used in ExoScreen assay and In vitro opsonization assay. CD9 antibody was used in in vivo metastasis assay and mass spectrometric immunoassay with monolith pipette tips. CD63 antibody was used in Membrane fusion assay.

User Reports

Examples of monoclonal antibodies recognizing exosome markers
Immunoelectron microscopy of exosomes using anti-exosome antibodies
Fluorescent immunostaining of exosomes produced by cancer cells using CD63 antibody (clone: 8A12)
Detection of exosomal proteins by Western blotting method using anti-CD81 antibody

Product List

Anti-Human Tetraspanin Monoclonal Antibodies

Product nameCatalog Number
Anti CD9, Human (Mouse) Unlabeled, 12A12 SHI-EXO-M01
Anti CD63, Human (Mouse) Unlabeled, 8A12 SHI-EXO-M02
Anti CD81, Human (Mouse) Unlabeled, 12C4 SHI-EXO-M03

Fluorescent-labeled Anti-Human Tetraspanin Monoclonal Antibodies

Tide Fluor™ 2 (TF2) has spectral properties similar to those of fluorescein and Alexa Fluor® 488 (Invitrogen). Compared to fluorescein, TF2 has much stronger fluorescence and higher photostability. Additionally, its fluorescence is pH-independent from pH 3 to 11. TF2-labeled antibodies exhibit much stronger fluorescence and higher photostability than those labeled with FITC. These characteristics make TF2 a superior alternative to fluorescein.
Tide Fluor™ 5 (TF5) has spectral properties essentially identical to those of Cy5. Compared to Cy5, TF5 has much stronger fluorescence and higher photostability. Additionally, its fluorescence is pH-independent from pH 3 to 11. TF5-labeled antibodies exhibit much stronger fluorescence and higher photostability than those labeled with Cy5. These characteristics make TF5 a superior alternative to Cy5.

Product nameCatalog Number
Anti CD9, Human (Mouse) Label Tide Fluor™ 2WS, 12A12 SHI-EXO-M01-TF2
Anti CD9, Human (Mouse) Label Tide Fluor™ 5 WS, 12A12 SHI-EXO-M01-TF5
Anti CD63, Human (Mouse) Label Tide Fluor™ 2WS, 8A12 SHI-EXO-M02-TF2
Anti CD63, Human (Mouse) Label Tide Fluor™ 5WS, 8A12 SHI-EXO-M02-TF5
Anti CD81, Human (Mouse) Label Tide Fluor™ 2WS, 12C4 SHI-EXO-M03-TF2
Anti CD81, Human (Mouse) Label Tide Fluor™ 5WS, 12C4 SHI-EXO-M03-TF5

Biotin-labeled Anti-Human Tetraspanin Monoclonal Antibodies

Product nameCatalog Number
Anti CD9, Human (Mouse) Biotin, 12A12 SHI-EXO-M01-B
Anti CD63, Human (Mouse) Biotin, 8A12 SHI-EXO-M02-B
Anti CD81, Human (Mouse) Biotin, 12C4 SHI-EXO-M03-B

Citations

■CD9 antibody
  1. S Tsuda et al., Sci Rep. 2017 Oct 11;7(1):12989. doi: 10.1038/s41598-017-13154-0.
  2. N Nishida-Aoki et al., Mol Ther. 2017 Jan 4;25(1):181-191. doi: 10.1016/j.ymthe.2016.10.009.
  3. K Matsuzaki et al., Oncotarget. 2017 Apr 11; 8(15): 24668-24678.　doi:10.18632/oncotarget.14969
  4. Kazutoshi Fujita et al., Sci Rep. 2017; 7: 42961. doi: 10.1038/srep42961
  5. Yoshioka Y et al., Nat Commun. 2014 Apr 7;5:3591. doi: 10.1038/ncomms4591.
  6. Saito S et al., Sci Rep. 2018 Mar 5;8(1):3997. doi: 10.1038/s41598-018-22450-2.
  7. Yagi Y et al., Neurosci Lett. 2017 Jan 1;636:48-57. doi: 10.1016/j.neulet.2016.10.042. Epub 2016 Oct 22.
  8. Ueda K et al., Sci Rep. 2014 Aug 29;4:6232. doi: 10.1038/srep06232.
  9. Tokuoka, S. M. et al. Lipid Profiles of Human Serum Fractions Enhanced with CD9 Antibody-Immobilized Magnetic Beads. Metabolites 12, 230 (2022).
■CD63 antibody
  1. N Nishida-Aoki et al., Mol Ther. 2017 Jan 4;25(1):181-191. doi: 10.1016/j.ymthe.2016.10.009.
  2. Yoshioka Y et al., Nat Commun. 2014 Apr 7;5:3591. doi: 10.1038/ncomms4591.
  3. Saito S et al., Sci Rep. 2018 Mar 5;8(1):3997. doi: 10.1038/s41598-018-22450-2.
■CD81 antibody
  1. M Somiya et al., J Extracell Vesicles. 2018 Feb 21;7(1):1440132. doi:10.1080/20013078.2018.1440132. eCollection 2018.
  2. Takahashi A et al., Nat Commun. 2017 May 16;8:15287. doi: 10.1038/ncomms15287.