|Host/Isotoype||Mouse / IgG1,k|
|Applications||ELISA, WB, Flow|
Tspan-29; S5.7; Target of the antiproliferative antibody; Tetraspanin-28;
|Storage instructions||Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze/thaw cycles.|
|Storage buffer||PBS(pH 7.4), containing 0.09% sodium azide.|
|Note||FOR RESEARCH ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES|
4A126.96.36.199 reacts with the CD81, a target for anti-proliferative antigen (TAPA-1) with 26 kDa MW, which ia a member of the TM4SF tetraspanin family. CD81 is broadly expressed on hemapoietic cells and enothelial and epithelial cells, but absent from erythrocytes and platelets as well as neutrophils. CD81 play role as a member of CD19/CD21/Leu-13 signal transdiction complex. It also is reported that anti-TAPA-1 antibody induce protein tyrosine phosphorylation that is prevented by increased intercellular thiol levels.
Anti-CD81(4A188.8.131.52) can detect several kinds of cell lines exosome via inderect ELISA assay
(A) To ensure antigen quality, exosome from different cell lines were measured by nanoparticle tracking analysis (NTA). Histogram shows mean peak and particle concentration of three selected cell lines exosomes, breast cancer cell line (AU565, MCF7), and colorectal cancer cell line (Colo205).
(B) Each kind of exosome was coated overnight with 5μg, and stained for CD81 with 4A184.108.40.206 at 1/1000 dilution in indirect ELISA assay. This data showed the P/N ratio, which means 1st Ab +2nd Ab signal / 2nd Ab only signal, were very high. This data can ensure the signal is coming from 4A220.127.116.11, not from non-specific binding.
WB analysis of different cell lines exosome using anti-CD81(4A18.104.22.168)
All lanes: Anti-CD81 antibody (4A22.214.171.124) at 1/1,000 dilution
Lane 1: Colo 205 cell line exosome (5μg)
Lane 2: MDA-MB-231 cell line exosomes (5μg)
Lane 3: MCF7 cell line exosomes (5μg)
Lane 4: DLD-1 cell lien exosome (5μg)
All lanes: Goat Anti-Mouse IgG1 at 1/20,000 dilution
Predicted band size: 25 kDa
Non-reducing form: sample treated with 4x Laemmli Sample Buffer
Plasma samples were fractionated by SEC (size exclusion chromatography) method. Exosome particles were predicted to exist at fractions 4-7, and the protein to exist in after the tenth tube in soluble form.
These results showed 4A126.96.36.199 antibodies could precisely capture exosome particles in fractions 4-7. On the other side, another brand CD81 antibody did not focus on exosome particles (still has high signal in fractions 10-16), which may cause non-specific signals.