Lumio™ Red In-Cell Detection Kit - FAQs

Yes, fluorescent protein-expressing cells can be fixed using 4% paraformaldehyde in PBS for 10 min followed by one quick PBS rinse and 3 x 5 min washes with 1 mL PBS.

Yes, all of the fluorescent proteins offered by (EmGFP, YFP, CFP, BFP and Cycle 3 GFP) have been humanized for optimal mammalian expression.

The 97 Da molecular weight change is due to the Enhancer capping the cysteines that aren't part of the Lumio tag to prevent nonspecific binding of the dye to the cysteines. So all cysteines except the 4 in the Lumio tag will be modified and show this change in molecular weight.

Lumio technology has at least two advantages over GFP: First, Lumio fluorescence can be detected both in vivo and in vitro, but GFP fluorescence can only be detected within the cell (in vivo) because proper protein folding is needed. Secondly, the Lumio tag is very small, only several amino acids long, which makes it very unlikely to affect the function of protein to which it is fused, whereas GFP is a large protein that may interfere with the function of the protein to which it is fused. The small size of Lumio tag can also useful when expressing short ORFs or peptides.