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Q1 – My GFP or RFP is only fluorescing weakly result in poor images, can NanoBoosters help?
Yes, Nano-Boosters bind specifically to your GFP or RFP fusion protein which allows the coupled fluorescent dye to dramatically increase the signal intensity and duration. For certain applications in which residual fluoresncence is present, we recommend matching the fluorescence emmision spectra of your GFP/RFP-fusion with the Nano-Booster as follows:
YFPs, GFPs and Citrin = GBA488
RFP mCherry = RBA594
mKate and mPlum = RBA647N
Q2 – My GFP or RFP fluorescence is completely bleached out before I can retrieve data. Can Nano-Boosters work in these cases?
Yes, Nano-Boosters are made to work after harsh fixation procedures that denature GFP and RFP fusions resulting in a loss of fluoresnce signal post-fixing. Also, because the set of ATTO dyes used is so much more robust than the GFP and RFP fluorophores, strong intensity excitation, like that of super resolution microscopy, work fine. In all of these cases, because the GFP and RFP endogenous signal is lost, you are free to select the ATTO dye most approriate for your experiment. We offer three choices linked to each of the NanoBooster Affinity Reagents: ATTO488, ATTO594 and ATTO647N.
Q3 – Which GFP variants does the GFP-Trap/ GFP-Booster recognize?
eGFP, wtGFP, GFP S65T
TagGFP
eYFP, YFP, Venus, Citrin
CFP
Not recognized:
TurboGFP
all RFPs
Q4 – Which RFP variants does the RFP-Trap/ RFP-Booster recognize?
mRFP
mCherry
mOrange
mPlum
mRFPruby
mKate2
Not recognized:
DsRed
TagRFP
all GFPs