Immunostaining of PFA fixed 3T3 cells expressing a TOM70-BFP reporter protein with FluoTag®-Q Abberior STAR 635P anti-TagFP sdAb (Cat. No. N0501, dilution 1:500, the BFP signal is represented in blue, the corresponding FluoTag®-signal is represented in yellow and the merge of both channels is represented in white).

FluoTag-Q anti-TagFP AF647

PFA-fixed Cos7 cells expressing a TOM70-BFP fusion protein were stained with FluoTag®-Q anti-TagFP coupled to Alexa Fluor 647 (Cat. No. N0501-AF647, dilution 1:500). A Greyscale image of the staining performed with N0501-AF647. B False color representation of the image shown in A is displayed in magenta (coloring according to the excitation wavelength of the employed fluorophore). C The corresponding DAPI signal of the depicted section. D Merge of A and C. False color representation of A in magenta and C in blue.

FluoTag-Q anti-TagFP AF568

PFA-fixed Cos7 cells expressing a TOM70-BFP fusion protein were stained with FluoTag®-Q anti- TagFP coupled to AZDye 568 (Cat. No. N0501-AF568, dilution 1:500). A Greyscale image of the staining performed with N0501-AF568. B False color representation of the image shown in A is displayed in red (coloring according to the excitation wavelength of the employed fluorophore). C The corresponding DAPI signal of the depicted section. D Merge of A and C. False color representation of A in red and C in blue.

FluoTag-Q anti-TagFP At488

PFA-fixed Cos7 cells expressing a TOM70-BFP fusion protein were stained with FluoTag®-Q anti- TagFP coupled to Atto 488 (Cat. No. N0501-At488, dilution 1:500). A Greyscale image of the staining performed with N0501-At488. B False color representation of the image shown in A is displayed in green (coloring according to the excitation wavelength of the employed fluorophore). C The corresponding DAPI signal of the depicted section. D Merge of A and C. False color representation of A in green and C in blue.

FluoTag®-Q anti-TagFP

Cat No: N0501 Category:

400,00 

FluoTag®-Q anti-TagFP is derived from an in-house developed single-domain antibody (sdAb) that recognizes fluorescent proteins deriving from Entacmaea quadricolor, such as mTagBFP, mTagRFP, mTagRFP657, and mKate with high affinity and specificity.

Read more

TagRFP was engineered in 2007 from TurboRFP (derived from Entacmaea quadricolor). Tag RFP displayed an extended fluorescence lifetime and showed robust photostability. In 2008 after further mutagenesis and the exchange of ~12 amino acid residues, this red-emitting protein turned into a blue-emitting protein TagBFP. Only several years later, in 2011 a monomeric version of TagBFP was described: mTagBFP2. 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 sdAb binds strongly to TagRFP, Tag BFP, and mTagBFP2. Look at our specificity chart in our 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.

Variations:
Conjugation Amount Cat No. RRID
Atto488 200 μl N0501-At488-L AB_2744611
AZDye568 200 μl N0501-AF568-L AB_3075930
Atto643 200 μl N0501-At643-L AB_3075933
Alexa647 200 μl N0501-AF647-L AB_3075931
AbberiorStar635P 200 μl N0501-Ab635P-L AB_2744610
Related Products: -
Clone: 1H7
Host: Alpaca
Produced in: E.coli
Application: IF
Dilution: 1:1000 (corresponding to 5 nM final concentration)
Capacity: N/A
Antigen: -
Targets: tagBFP
Specificity: Recognizes TagRFP, Tag BFP and mTagBFP2. Probably also other derivatives not yet tested.
Formulation: A 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 in 200 µl, the concentration of single-domain antibody is 5 µM
kDa: -
Ext Coef: -
Shipping: Ambient temperature
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.
Protocols:

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.

 

References:
  1. Oleksiievets N, Sargsyan Y, Thiele JC, et al. Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells. Commun Biol. 2022;5(1):38. Published 2022 Jan 11. doi:10.1038/s42003-021-02976-4 (FL-DNA PAINT)
  2. Thiele JC, Helmerich DA, Oleksiievets N, et al. Confocal Fluorescence-Lifetime Single-Molecule Localization Microscopy. ACS Nano. 2020;14(10):14190-14200. doi:10.1021/acsnano.0c07322 (DNA PAINT)
  3. Sograte-Idrissi S, Oleksiievets N, Isbaner S, et al. Nanobody Detection of Standard Fluorescent Proteins Enables Multi-Target DNA-PAINT with High Resolution and Minimal Displacement Errors. Cells. 2019;8(1):48. Published 2019 Jan 14. doi:10.3390/cells8010048 (DNA PAINT)
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.