Anti CD9, Rat Monoclonal Antibody(77B), Biotin-conjugated
- for Immunochemistry
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at -20 degrees C.
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50uL
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In stock in Japan |
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Product Information
| Cross Reactivity | Human [Note] This antibody doesn’t recognize the mouse and bovine CD9. It shows a little reactivity with rat CD9. |
|---|---|
| Clone No. | 77B |
| Host | Rat |
| Subclass | IgG2b |
| Formulation | 1xTBS aqueous solution with 50w/v% glycerol, 0.05w/v% sodium azide. |
| Label | Biotin |
▼Evaluating the reactivity
The reactivity of this antibody against a serum of each organisms was measured by ELISA.
When the signal value was the same level as the blank, it was judged as “non-reactive”.
Application
| ELISA | 1:2,000 – 1:16,000 |
|---|---|
| Western Blot | 1:1,000 – 1:8,000 * Please use this antibody under non-reducing condition |
| Immunoprecipitation | 1:100 – 1:1,000 |
Please optimize the most appropriate concentration for your analysis and samples.
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].
| Sample | Exosomes from cell culture supernatant of COLO201 cells |
|---|---|
| ELISA Kit | PS Capture™ Exosome ELISA Kit (Streptavidin HRP) [code No. 298-80601] |
| Detection antibody | 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] (250 ng/mL) 30B: Anti CD9, Rat Monoclonal Antibody (30B), Biotin-conjugated [code No. 019-27953] (250 ng/mL) |
| 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. |
▶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.
| Sample | Exosomes from cell culture supernatant of COLO201 cells |
|---|---|
| Immobilized antibody | Anti CD9 Ab: Anti CD9, Rat Monoclonal Antibody (77B) [code No. 019-28173] Anti CD63 Ab: Anti CD63, Monoclonal Antibody (3-13) [code No. 012-27063] Anti CD81 Ab: Anti CD81, Monoclonal Antibody (17B1) [code No. 011-27773] |
| Detection antibody | 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] (250 ng/mL) 30B: Anti CD9, Rat Monoclonal Antibody (30B), Biotin-conjugated [code No. 019-27953] (250 ng/mL) |
| Detection | HRP-conjugated Streptavidin and TMB solution |
| 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. |
ELISA
Human serum and human heparin plasma were detected by PS ELISA with this antibody.
| Sample | Human serum and Human heparin plasma [Step Dilution] |
|---|---|
| ELISA Kit | PS Capture™ Exosome ELISA Kit (Streptavidin HRP) [code No. 298-80601] |
| Detection antibody | 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 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 [code No. 017-28211] (1 μg/mL) |
| Detection reagent | HRP-Streptavidin, ImmunoStar® Zeta [code No. 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).
| Immunoprecipitation | |
|---|---|
| Sample | Cell culture supernatant of HEK293T cells |
| Capture antibody | Rat IgG: Biotinylated anti-Rat IgG antibody (as negative control) 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] |
| 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 [code No. 291-72401 |
| 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. |
Overview / Applications
| Outline | CD9 is a cell surface glycoprotein which belongs to the tetraspanin superfamily (TM4SF) and, is found on the surface of exosomes. This is known to form a complex with integrins and other TM4SF, can modulate cell adhesion and migration. Also, this induces platelet activation and aggregation. CD9 is known as exosome marker protein as well as CD63 and CD81. This antibody is established against human CD9 with Rat, and conjugated with biotin. It specifically recognizes human CD9 and very slightly rat CD9. It does not recognize bovine and mouse CD9. [Clone No.] 77B [Immunized animal] Rat [Antibody subclass] IgG2b [Label] Biotin |
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| Application | ELISA 1:2000 - 1:16000 Western Blot 1:1000 - 1:8000 *Use this antibody under non-reducing condition. Immunoprecipitation 1:100 - 1:1000 Please optimize the most appropriate concentration for your analysis and samples. |
Property
| Specificity | Human CD9. Note: It shows a little reactivity with rat CD9. It does not react with bovine and mouse CD9. [Reactivity confirmation condition] The reactivity of this antibody against the serum of each organisms was measured by ELISA. When the signal value was the same level as the blank, it was judged as "not react". |
|---|---|
| Composition | 1xTBS, 50w/v% glycerol and 0.05% sodium azide. |
| Concentration | antibody concentration : Indicated on the label. |
Manufacturer Information
Alias
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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