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What positive control for intracellular staining?

(flow cytometry)

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#1 Paja

Paja

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Posted 28 November 2011 - 11:21 AM

Hi,
I need to learn well technique of intracellular staining of PBMC on flow cytometry in order to make sure that later, when I do not see protein of my interest, the reason is that this protein is not expressed by the cell not because I am not able to make the procedure the correct way. But, I do not know what primary antibody, against what protein to buy and use. What intracellular protein I shall use to train detection by FCM to be 100% positive it is always expressed in PBMC...?

Thank you!
Paja

#2 RynDggn

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Posted 28 November 2011 - 07:31 PM

When people need to ensure their cells are properly permeabilized and in good condition, they'll sometimes use an antibody to F-Actin, which is present in all cells. If the F-Actin staining is always positive then at least you know that the cells are properly permeabilized and that your buffers are working properly. This could serve as a positive control.




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