I want to do these two tests but I can not understand my protocol. Is there anyone to help me to learn how to do it.
Thank you so much in advance for your help.
Karen A. Darbinyan
Hello, Sahar Ebrahimi Samani
TRAP staining and quantifying below:
Aspirate medium and wash cells with 200µl PBS. Aspirate PBS.
Add 100µl Citrate buffer + 0.1% Triton X-100, incubate 5 minutes @RT.
Add 100µl pNPP solution, incubate 5 minutes @RT.
Stop reaction by adding 50µl 0.5M NaOH.
Read OD at 405nm.
pNPP solution:
pNPP - 40mg
Tartrate solution - 1.8ml
Citrate buffer - 5.7ml
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Sahar Ebrahimi Samani
Thank you so much for your help.
What about the supernatant that we can use for quantification?
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