GFP and RFP-Booster for better immunofluorescence imaging


What are Nano-Booster?

Nano-Booster  are fluorescent probes that enhance the signal of fluorescent proteins. They are very small (15kDa) and consist of an antibody fragment conjugated to a fluorescent dye. The antibody fragment is derived from antigen binding domain of  heavy chain only antibody and is called VHH or Nanobody (you can find more info about Nanobodies  here). The GFP-Booster is an anti-GFP Nanobody that binds to CFP, GFP, AcGFP, mClover (Clover A206K), YFP, etc., and the RFP-Booster is an anti-RFP Nanobody that binds to mRFP, mCherry, mRFPruby, etc. Here you can find a complete list of tested fluorescent proteins:  Fluorescent protein specificity table Nano-Booster.  The GFP- and RFP-Booster are optimized for immunofluorescence.

ChromoTek's GFP- and RFP-Booster are compatible with conventional antibodies and  Nano-Secondaries  for multiplexing. GFP-Booster  and  RFP-Booster  conjugated to Alexa Fluor® 488, 568, and 647:

  • Stronger signal in confocal and standard fluorescence microscopy
  • More options in super-resolution microscopy including STORM and MINFLUX
  • Constant degree of labeling (DOL = 2 dyes per Booster) for higher resolution
  • More than 95% labeling efficiency

 

Final_DAPI_TOM70GFP_gbaf488_191114_csl6-2

Confocal image of HeLa cells transiently transfected with Tom70-eGFP and immunostained with GFP-Booster Alexa Fluor 488 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 µm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

 

 

Available Alexa Fluor conjugates for more flexibility:

  • GFP-Booster  conjugated to Alexa Fluor 488, Alexa Fluor® 568, and Alexa Fluor® 647
  • RFP-Booster  conjugated to Alexa Fluor 568 and Alexa Fluor® 647

alexa fluor emission wavelength

 

 

Here is an example for multiplexing with GFP-Booster Alexa Fluor 488 and RFP-Booster Alexa Fluor 568.

Option_gb2af488_rb2af568

HeLa cells transiently transfected with PCNA-mRFP and Tom70-eGFP were subjected to one-step immunostaining with RFP-Booster Alexa Fluor 568 (red) and GFP-Booster Alexa Fluor 488 (green). Scale bar, 10 µm. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

 

 

GFP- and RFP-Booster are anti-GFP and anti-RFP Nanobodies conjugated to fluorescent dyes. They stabilize, enhance and reactivate GFP and RFP signals: