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Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated Application Data
Characteristic comparison with Clone No. 30B by ELISA
The characteristics of this product and our another anti CD9 antibody were compared by ELISA.
The two type of antibodies (77B and 30B) were compared in PS ELISA [our original ELISA kit with Tim4 protein immobilized].
Both the signal value and the S/B ratio of 30B were higher. It has been shown that it is suitable to use 30B as a detection antibody in PS ELISA.
Sam
| Samples | Exosomes from cell culture supernatant of COLO201 cells |
| Elisa Kit | PS Capture Exosome ELISA Kit (Streptavidin HRP) (Catalog ID 298-80601) |
| Detection antibody | 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated (Catalog ID 017-28211) (250 ng/mL) 30B: Anti CD9, Rat Monoclonal Antibody (30B), Biotin-conjugated (Catalog ID 019-27953) (250 ng/mL) |
Two types of anti-CD9 antibodies (77B and 30B) were used for the immobilization antibody and the detection antibody in the antibody-immobilized ELISA for comparison.
Clone No. 30B had higher signal and blank value in antibody-immobilized ELISA, when 30B was used as the detection antibody.
Clone No. 77B had higher S/N ratio than clone No. 30B when compared in terms of S/N ratio.
It has been shown that it is suitable to use 77B as a detection antibody in antibody-immobilized ELISA.
Sam
| Samples | Exosomes from cell culture supernatant of COLO201 cells |
| Immobilied antibody | Anti CD9 Ab: Anti CD9, Rat Monoclonal Antibody (77B) (Catalog ID 019-28173) Anti CD63 Ab: Anti CD63, Monoclonal Antibody (3-13) (Catalog ID 012-27063) Anti CD81 Ab: Anti CD81, Monoclonal Antibody (17B1) (Catalog ID 011-27773) |
| Detection antibody | 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated (Catalog ID 017-28211) (250 ng/mL) 30B: Anti CD9, Rat Monoclonal Antibody (30B), Biotin-conjugated (Catalog ID 019-27953) (250 ng/mL) |
| Detection | HRP-conjugated Streptavidin and TMB solution |
ELISA
Human serum and human heparin plasma were detected by PS ELISA with this antibody.
It was shown that the signal value fluctuated depending on the dilution ratio, and Exosomes in human serum and heparin plasma could be detected.
| Sample | Human serum and Human heparin plasma [Step Dilution] |
| ELISA Kit | PS Capture Exosome ELISA Kit (Streptavidin HRP) (Catalog ID 298-80601) |
| Detection antibody | 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated (Catalog ID 017-28211) (250 ng/mL) |
It was shown that the signal value fluctuated depending on the dilution ratio, and Exosomes in human serum and heparin plasma could be detected.
Western Blot
Isolated exosomes derived from culture supernatant, human serum, and human heparin plasma were electrophoresed under non-reducing conditions, and Western Blot was performed using this antibody.
| Sample | Exosomes isolated from cell culture supernatant of COLO201 cells, human serum, and human heparin plasma. |
| Detection antibody | Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated (Catalog ID 017-28211) (1 μg/mL) |
| Detection reagent | HRP-Streptavidin, Immunostar Zeta (Catalog ID 291-72401) |
CD9 protein (24kDa) in each sample (non-reducing condition) was detected with this antibody.
Immunoprecipitation
The culture supernatant was immunoprecipitated with Anti Rat IgG antibody and this antibody.
Immunoprecipitated samples were analyzed by Western Blot with Anti-CD9 antibody and Anti-Alix antibody (competitors’ products).
CD9 and Alix which are marker of exosomes were detected in the sample. immunoprecifitated with this antibody.
It was shown that exosomes in samples can be selectively captured by using this product for immunoprecipitation.
| Immunoprecipitation | |
| Sample | Cell culture supernatant of HEK293T cells |
| Detection antibody |
Rat IgG: Biotinylated anti-Rat IgG antibody (as negative control) 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated (Catalog ID 017-28211) (1 μg/mL) |
| Immunoprecipitated beads | Streptavidin-conjugated magnetic beads |
| Western Blot | |
| Primary antibody | CD9: Anti-CD9 antibody (competitor’s) [1 μg/mL] Alix: Anti-Alix antibody (competitor’s) [1 μg/mL] |
| Secondary antibody |
HRP-conjugated Anti-Mouse IgG antibody |
| Detection reagent | Immunostar Zeta (Catalog ID 291-72401) |
