Immunostaining of PFA fixed 3T3 cells expressing a TOM70-GFP reporter protein with a FluoTag®-Q Atto 647N anti-GFP sdAb (Cat. No. N0301-At647N, dilution 1:500, the GFP signal is represented in green, the corresponding FluoTag®-signal is represented in red and the merge of both channels is represented in yellow). Nuclei were visualized by DAPI staining (blue).
FluoTag®-Q anti-GFP is derived from an in-house developed single-domain antibody (sdAb) that recognizes GFP and its most common derivatives with high affinity and specificity.
A breakthrough in biology and bioluminescence began in the 60s with the discovery of a glowing protein obtained from the jellyfish Aequorea victoria by Osamu Shimomura and colleagues. However, it was only in the early 90s when the green fluorescent protein (GFP) sequence was cloned and used inside a foreign organism as a fluorescent marker. Today more than 800 entries of various fluorescent proteins can be found in an open-source database with the most currently available variants in the fluorescence protein database “fpbase”
Our sdAbs bind strongly to most of the fluorescent proteins derived from Aequorea Victoria; check our specificity chart in the Resource Section.
FluoTags® can be equipped with a single fluorophore for more quantitative readouts (FluoTag-Q), with two fluorophores per single-domain antibody (FluoTag-X2), and we also developed a blend of two sdAbs bindings simultaneously the target proteins and each bearing two fluorophores (FluoTag-X4). For more detailed information on the FluoTags, please check our Technology Section.
|Produced in:||E. coli|
|Dilution:||1:1000 (corresponding to 5 nM final concentration)|
|Specificity:||Recognizes GFP (green fluorescent protein) and common GFP derivatives like EGFP, mEGFP, Sirius, tSapphire, Cerulean, eCFP, mTurquoise, acGFP, Emerald, superecliptic pHluorin, paGFP, superfolder GFP, eYFP, mVenus and Citrine. Probably also other derivatives not yet tested.|
|Formulation:||A single sdAb clone was lyophilized from PBS pH 7.4. Reconstitute with 200 µL of 50 % glycerol in deionized water. We recommend including 0.1 % sodium azide as a preservative if applicable. When reconstituted in 200 µl, the concentration of single-domain antibody is 5 µM|
|Storing:||Vials containing lyophilized protein can be stored at 4 °C for 6 months. We recommend reconstituting the protein with 50 % sterile glycerol in water including 0.1 % sodium azide as preservative if applicable. Minimize the number of freeze-thaw cycles by aliquoting the reconstituted protein. Long term storage at -80 °C for up to 6 months. Working aliquots can be stored at -20 °C for up to 4 weeks. We do not recommend storing the reconstituted protein at 4 °C.|
Western Blotting is not recommended with this product, sdAbs tend to recognize native protein conformation only.
Look at detailed protocols and our specificity chart in our Resource Section.
|Notice:||To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.|
|Legal terms:||By purchasing this product you agree to our general terms and conditions.|