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Team:TU Darmstadt/Protocols/Antibody staining
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<li><a href="/Team:TU_Darmstadt/Safety" title="Safety">Safety</a></li> | <li><a href="/Team:TU_Darmstadt/Safety" title="Safety">Safety</a></li> | ||
<li><a href="/Team:TU_Darmstadt/Downloads" title="Downloads">Downloads</a></li></ul></li> | <li><a href="/Team:TU_Darmstadt/Downloads" title="Downloads">Downloads</a></li></ul></li> | ||
| - | <li><a href="/Team:TU_Darmstadt/Human_Practice" title="Human Practice">Human Practice</a | + | <li><a href="/Team:TU_Darmstadt/Human_Practice" title="Human Practice">Human Practice</a></li> |
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| - | + | ||
| - | + | ||
<li><a href="/Team:TU_Darmstadt/Sponsors" title="Sponsors">Sponsors</a><ul> | <li><a href="/Team:TU_Darmstadt/Sponsors" title="Sponsors">Sponsors</a><ul> | ||
<li><a href="/Team:TU_Darmstadt/Sponsors" title="Sponsors">Overview</a></li> | <li><a href="/Team:TU_Darmstadt/Sponsors" title="Sponsors">Overview</a></li> | ||
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* 1. antibody (mouse-anti-myc) | * 1. antibody (mouse-anti-myc) | ||
* 2. antibody (goat-antimouse-biotin) | * 2. antibody (goat-antimouse-biotin) | ||
| - | * SAPE | + | * Streptavidin, R-phycoerythrin conjugate (SAPE) |
== Procedure == | == Procedure == | ||
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# incubate on ice for 10 min. | # incubate on ice for 10 min. | ||
# wash two times with 200 µL PBS buffer | # wash two times with 200 µL PBS buffer | ||
| - | * now your cells are ready to be detected | + | * now your cells are ready to be detected via flow cytometry |
Latest revision as of 21:19, 26 September 2012
Antibody staining
Materials
- ice bath
- Centrifuge
- PBS buffer pH 7.4
- 1. antibody (mouse-anti-myc)
- 2. antibody (goat-antimouse-biotin)
- Streptavidin, R-phycoerythrin conjugate (SAPE)
Procedure
- centrifuge 750 µL of cell suspension for 2 min in order to gain pellet
- wash the pellet two times with 200 µL PBS buffer (after every wash centrifuge for 2 min)
- add 30 µL of 1. antibody 1:10 concentration in PBS buffer
- incubate suspension including antibody for 10 min. on ice
- centrifuge for 2 min.
- wash pellet three times with 200 µL PBS buffer (after every wash centrifuge for 2 min)
- add 10 µL of 2. antibody 1:10 concentration in PBS buffer
- incubate suspension including antibody for at least 30 min on ice
- centrifuge for 2 min
- wash pellet three times with 200 µL PBS buffer (after every wash centrifuge for 2 min)
- add 10 µL of SAPE 1:10 concentration in PBS buffer
- incubate on ice for 10 min.
- wash two times with 200 µL PBS buffer
- now your cells are ready to be detected via flow cytometry
