Immunostaining of PFA fixed Cos7 cells expressing a TOM70-mRuby reporter protein with FluoTag®-X2 Alexa Fluor 647 anti-mRuby sdAb (Cat. No. N3302-AF647-L, dilution 1:500, the mRuby signal is represented in red, the corresponding FluoTag-signal is represented in green and the merge of both channels is represented in yellow). Nuclei were visualized by DAPI staining (blue).
FluoTag®-X2 anti-mRUBY is derived from an in-house developed single-domain antibody (sdAb) that recognizes mRuby and mRuby derivatives, which is a bright monomeric red fluorescent protein derived from Entacmaea quadricolar.
mRuby is a bright monomeric red fluorescent protein, which has more than 20 mutations compared to its wild-type predecessor (eqFP611) found in Entacmaea quadricolor . mRuby, mRuby2 & mRuby3 are all exited at 560 nm, with mRuby3 being the brightest.
An open-source database with the most currently available variants can be found in the fluorescence protein database “fpbase”
Our nanobody binds specifically and strongly to mRuby fluorescent proteins. Have a look at our specificity chart in our Resource Section.
FluoTags are directly conjugated to fluorophores, however, they 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, decorating the target protein with 4 fluorophores in total (FluoTag-X4). For more detailed information on the FluoTags, please check our Technology Section.
|Dilution:||1:500 (corresponding to 5 nM final concentration)|
|Specificity:||Recognizes most common mRuby derivatives|
|Formulation:||The single sdAb clone was lyophilized from PBS pH 7.4, containing 2% BSA (US-Origin). Reconstitute with 200 µL of 50 % glycerol in deionized water. We recommend including 0.1 % sodium azide as a preservative if applicable. When reconstituted with 200 µl, the concentration of single-domain antibody is 2.5 µM|
|Storing:||Vials containing lyophilized protein can be stored at 4 °C for 6 months. We recommend reconstituting the protein with 50 % sterile glycerol 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. The 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.|