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 1o Antibody 2o 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 oAb
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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Anti-GRP94 Antibody (PA1340)

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