Mini Guide to FACS Staining Controls

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BosterBio | Antibodies, ELISA Kits, Detection Kits
March 31, 2017
Dr. Booster

Dear Earthling,

With great versatility (in FACS panels and protocols), comes great responsibility in choosing the right controls. Take a quick look to make sure you have included all the appropriate ones! Here’s a convenient mini guide to FACS staining controls:

Control 1 o Antibody 2 o Antibody Why use it?
Unstained + + To subtract autofluorescence
Unstimulated +/− +/− To set baseline cytokine/phosphoprotein expression
1 o /2 o Ab treated negative cells +/− +/− To check for cross-reactivity of the antibodies
Isotype Only isotype + for indirect staining To subtract non-specific Fc receptor binding of 1 o Ab
Compensation for every fluorochrome + + for indirect staining To subtract spectral overlap/multichannel spillage
FMO for every fluorochrome + + for indirect staining To subtract the signal of all non-specific fluorochromes from a specific channel
Viability - - Live/dead cell discrimination
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Dr. Booster BosterBio | Antibodies, ELISA Kits, Detection Kits

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