Immunostaining of PFA fixed 3T3 cells expressing a TOM70-BFP reporter protein with FluoTag®-X2 Abberior STAR 580 anti-TagFP sdAb (Cat. No. N0502, dilution 1:500). The BFP signal is represented in blue, the corresponding FluoTag®-signal is represented in red and the merge of both channels is represented in magenta.

FluoTag-X2 anti-TagFP AF647

PFA-fixed Cos7 cells expressing a TOM70-BFP fusion protein were stained with FluoTag®-X2 anti-TagFP coupled to Alexa Fluor 647 (Cat. No. N0502-AF647, dilution 1:500). A Greyscale image of the staining performed with N0502-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-X2 anti-TagFP AF568

PFA-fixed Cos7 cells expressing a TOM70-BFP fusion protein were stained with FluoTag®-X2 anti-TagFP coupled to AZDye 568 (Cat. No. N0502-AF568, dilution 1:500). A Greyscale image of the staining performed with N0502-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-X2 anti-TagFP At488

PFA-fixed Cos7 cells expressing a TOM70-BFP fusion protein were stained with FluoTag®-X2 anti-TagFP coupled to Atto 488 (Cat. No. N0502-At488, dilution 1:500). A Greyscale image of the staining performed with N0502-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®-X2 anti-TagFP

Cat No: N0502 Category:

400,00 

FluoTag®-X2 anti-TagFP is an in-house developed single-domain antibody (sdAb) with high affinity and specificity for fluorescent proteins derived from Entacmaea quadricolor, including TagRFP, TagRFP657, TagBFP, mTagBFP2, mKate, and mKate2. It features two fluorophores per sdAb molecule (X2).

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Fluorescent proteins (FPs), widely used as protein tags in cell biology and microscopy, have revolutionized protein imaging and biochemical applications. TagRFP, a monomeric red fluorescent protein, was engineered from TurboRFP (derived from Entacmaea quadricolor) to exhibit an extended fluorescence lifetime and enhanced photostability (Merzlyak et al., 2007). Through further mutagenesis, this red-emitting protein was transformed into the blue-emitting TagBFP (Subach et al., 2008). A few years later, an improved version known as mTagBFP2 was introduced, offering greater brightness and stability (Subach et al., 2011). In 2007, mKate was developed from TagRFP, noted for its excellent far-red fluorescence and photostability (Shcherbo et al., 2007). The most currently available variants can be found on an open-source Fluorescent Protein Database FPbase.

FluoTag®-X2 anti-TagFP binds strongly to TagRFP, TagRFP657, TagBFP, mTagBFP2, mKate and mKate2. For detailed information, please refer to our FP Specificity Chart.

FluoTags® are single-domain antibodies (sdAbs) directly conjugated to a defined number of fluorophores. FluoTag®-Q is sdAb equipped with a single fluorophore for more quantitative readouts. FluoTag®-X2 carries two fluorophores per sdAb. FluoTag®-X4 is a blend of two different sdAbs that bind simultaneously to the single target, each bearing two fluorophores, thereby decorating the target protein with 4 fluorophores in total for a brighter signal. For more detailed information on FluoTags®, please check our Technology Section.

MSDS
Variations:
Conjugation Amount Cat No. RRID
Atto488 200 μl N0502-At488-L AB_2744623
AZDye568 200 μl N0502-AF568-L AB_3075935
Atto643 200 μl N0502-At643-L AB_3075938
Alexa647 200 μl N0502-AF647-L AB_3075936
AbberiorStar635P 200 μl N0502-Ab635P-L AB_2744622
Related Products: -
Clone: 1H7
Host: Alpaca
Produced in: E.coli
Application: IF
Dilution: 1:500 (corresponding to 5 nM final concentration)
Capacity: N/A
Antigen: -
Targets: TagFP
Specificity: Recognizes TagRFP, TagRFP657, TagBFP, mTagBFP2, mKate and mKate2. Likely reactive with other untested TagRFP derivatives.
Formulation: The 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 final concentration of single-domain antibody is 2.5 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 deionized water and including 0.1% sodium azide as a 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:

These products are not recommended for detecting proteins in Western blot, as sdAbs primarily recognize native or folded proteins.

For more details check our Protocols in Resource Section and FP Specificity Chart.

References:
  1. Hutto RA, Rutter KM, Giarmarco MM, Parker ED, Chambers ZS, Brockerhoff SE. Cone photoreceptors transfer damaged mitochondria to Müller glia. Cell Rep. 2023;42(2):112115. doi:10.1016/j.celrep.2023.112115 (IHC, zebrafish)
  2. Pouchelon G, Vergara J, McMahon J, et al. A versatile viral toolkit for functional discovery in the nervous system. Cell Rep Methods. 2022;2(6):100225. Published 2022 May 26. doi:10.1016/j.crmeth.2022.100225
  3. Gombkoto P, Gielow M, Varsanyi P, Chavez C, Zaborszky L. Contribution of the basal forebrain to corticocortical network interactions. Brain Struct Funct. 2021;226(6):1803-1821. doi:10.1007/s00429-021-02290-z
  4. Pulido C, Ryan TA. Synaptic vesicle pools are a major hidden resting metabolic burden of nerve terminals. Sci Adv. 2021;7(49):eabi9027. doi:10.1126/sciadv.abi9027 (IF/ICC; rat neurons)
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 & Conditions.