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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 AF647

PFA-fixed Cos7 cells expressing a TOM70-nfGFP-BFP fusion protein (nf: non-fluorescent) were stained with FluoTag®-Q anti-GFP coupled to Alexa Fluor 647 (Cat. No. N0301-AF647, dilution 1:500). A Greyscale image of the staining performed with N0301-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 BFP 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-GFP AF568

PFA-fixed Cos7 cells expressing a TOM70-nfGFP-BFP fusion protein (nf: non-fluorescent) were stained with FluoTag®-Q anti-GFP coupled to AZDye 568 (Cat. No. N0301-AF568, dilution 1:500). A Greyscale image of the staining performed with N0301-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 BFP 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-GFP At488

PFA-fixed Cos7 cells expressing a TOM70-nfGFP-BFP fusion protein (nf: non-fluorescent) were stained with FluoTag®-Q anti-GFP coupled to Atto 488 (Cat. No. N0301-At488, dilution 1:500). A Greyscale image of the staining performed with N0301-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 BFP 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-GFP AF647
FluoTag-Q anti-GFP AF568
FluoTag-Q anti-GFP At488

FluoTag®-Q anti-GFP

Cat No: N0301 Category:

400,00 €

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.

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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.

MSDS Protocols
Variations:
Conjugation Amount Cat No. RRID
Atto488 200 μl N0301-At488-L AB_2744617
AZDye568 200 μl N0301-AF568-L AB_3075898
AbberiorStar635P 200 μl N0301-Ab635P-L AB_2744616
Atto643 200 μl N0301-At643-L AB_3075901
Alexa647 200 μl N0301-AF647-L AB_2905515
Related Products: -
Clone: 1H1
Host: Alpaca
Produced in: E. coli
Application: IF
Dilution: 1:1000 (corresponding to 5 nM final concentration)
Capacity: N/A
Antigen: -
Targets: GFP
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 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 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.
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:
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  12. Mookherjee D, Das S, Mukherjee R, et al. RETREG1/FAM134B mediated autophagosomal degradation of AMFR/GP78 and OPA1 -a dual organellar turnover mechanism. Autophagy. 2021;17(7):1729-1752. doi:10.1080/15548627.2020.1783118 (STED)
  13. Kutz S, Zehrer AC, Svetlitckii R, et al. An Efficient GUI-Based Clustering Software for Simulation and Bayesian Cluster Analysis of Single-Molecule Localization Microscopy Data. Front Bioinform. 2021;1:723915. Published 2021 Oct 11. doi:10.3389/fbinf.2021.723915 (dSTORM)
  14. Tejada-Arranz A, Galtier E, El Mortaji L, Turlin E, Ershov D, De Reuse H. The RNase J-Based RNA Degradosome Is Compartmentalized in the Gastric Pathogen Helicobacter pylori. mBio. 2020;11(5):e01173-20. Published 2020 Sep 15. doi:10.1128/mBio.01173-20 (SMLM)
  15. Mészáros J. Regulation of exocytosis and peptide release by synaptotagmins in sensory neurons. PhD thesis (2020), University of Sheffield.
  16. Hoff M. Combinatorial analysis of nanobody-detected epitopes for protein identification. PhD thesis (2020), University of Göttingen.
  17. Thevathasan JV, Kahnwald M, Cieśliński K, et al. Nuclear pores as versatile reference standards for quantitative superresolution microscopy [published correction appears in Nat Methods. 2019 Dec;16(12):1332]. Nat Methods. 2019;16(10):1045-1053. doi:10.1038/s41592-019-0574-9 (STED)
  18. Seitz KJ, Rizzoli SO. GFP nanobodies reveal recently-exocytosed pHluorin molecules. Sci Rep. 2019;9(1):7773. Published 2019 May 23. doi:10.1038/s41598-019-44262-8
  19. Wagner W, Lippmann K, Heisler FF, et al. Myosin VI Drives Clathrin-Mediated AMPA Receptor Endocytosis to Facilitate Cerebellar Long-Term Depression. Cell Rep. 2019;28(1):11-20.e9. doi:10.1016/j.celrep.2019.06.005 (STED)
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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.

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