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Species-independent multiplexing with Smart Secondaries® conjugated to abberior STAR dyes_FluoTag-X2 anti-Rabbit IgG_N2402

Multiplexing (see our Technologies): PFA-fixed mammalian cells were stained using four rabbit primary antibodies premixed with FluoTag®-X2 anti-Rabbit IgG (Smart Secondaries®) conjugated to different abberior STAR dyes. Peroxisomes stained with anti-PMP70 rabbit polyclonal Ab and FluoTag®-X2 anti-Rabbit IgG conjugated to abberior STAR 460L (cyan; Cat. No. N2402-Ab460L-S). Golgi apparatus stained with anti-Giantin rabbit polyclonal Ab and FluoTag®-X2 anti-Rabbit IgG conjugated to abberior STAR GREEN (green; Cat. No. N2402-AbGREEN-S). Nuclear pore complexes stained with anti-NUP98 rabbit monoclonal Ab and FluoTag®-X2 anti-Rabbit IgG conjugated to abberior STAR ORANGE (magenta; Cat. No. N2402-AbORANGE-S ). Mitochondria stained with anti-TOM20 rabbit polyclonal Ab and FluoTag®-X2 anti-Rabbit IgG conjugated to abberior STAR RED (orange; Cat. No. N2402-AbRED-S).

 Multiplexing with same-species primary antibodies and Smart Secondaries (two FluoTag®-X2 anti-Mouse IgG1 abberior STAR ORANGE & STAR RED) and FluoTag®-X4 anti-GFP abberior deepRED_NanoTag

Multiplexing (see our Technologies): PFA-fixed U2OS-NUP96-GFP stable cells were stained using two mouse primary antibodies, each pre-mixed with FluoTag®-X2 anti-Mouse IgG1 (Smart Secondaries®), and subsequently combined with FluoTag®-X4 anti-GFP. All FluoTag® reagents were conjugated to distinct abberior STAR dyes. NUP96-GFP was detected using FluoTag®-X4 anti-GFP conjugated to deepRED (A, red; Cat. No. N0304-AbdeepRED-L), which specifically binds to GFP fused to NUP96. Nuclear pore complexes (NPCs) were stained with NPC marker mouse monoclonal Ab and FluoTag®-X2 anti-Mouse IgG1 conjugated to abberior STAR RED (B, blue; Cat. No. N2002-AbRED-S). Vimentin was stained with anti-Vimentin mouse monoclonal Ab and FluoTag®-X2 anti-Mouse IgG1 conjugated to abberior STAR ORANGE (C, merged, orange; Cat. No. N2002-AbORANGE-S).

 N2402_Multiplexing_Smart Secondaries®_NanoTag

Multiplexing (see our Technologies): Indirect immunostaining of PFA fixed Cos7 cells with an anti-Erd2 rabbit polyclonal and affinity-purified primary antibody (Cat. No. 210 003, Synaptic Systems, dilution 1:500) detected with FluoTag®-X2 Atto 488 anti-Rabbit IgG (Cat. No. N2402-At488-S) and an anti-Aif rabbit polyclonal and affinity-purified primary antibody (Cat. No. 300 003, Synaptic Systems, dilution 1:500) detected with FluoTag®-X2 anti-rabbit IgG abberior STAR 580 (Cat. No. N2402-Ab580-S). Nuclei were visualized by DAPI staining (blue).
Species-independent multiplexing with Smart Secondaries® conjugated to abberior STAR dyes_FluoTag-X2 anti-Rabbit IgG_N2402
Multiplexing with same-species primary antibodies and Smart Secondaries (two FluoTag®-X2 anti-Mouse IgG1 abberior STAR ORANGE & STAR RED) and FluoTag®-X4 anti-GFP abberior deepRED_NanoTag
N2402_Multiplexing_Smart Secondaries®_NanoTag

FluoTag®-X2 anti-Rabbit IgG

Cat No: N2402 Category:

205,00 €

This new FluoTag®-X2 anti-Rabbit IgG (Smart Secondaries®) is a species-specific FluoTag® directed against IgGs from Rabbit. It serves as an effective secondary tool, enabling OneStep Immunofluorescence and species-independent Multiplexing.

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FluoTag®-X2 anti-Rabbit IgG is a fluorescent single-domain antibody (sdAb, nanobody, VHH) with high specificity and affinity for Rabbit IgG. It binds the constant Fc domain of any Rabbit IgG, regardless of clonality or VH domain, making it an ideal secondary tool for demanding imaging applications. As part of our Smart Secondaries® range, the FluoTag®-X2 anti-Rabbit IgG is particularly suited for OneStep Immunofluorescence (IF) and species-independent Multiplexing.

FluoTag®-X2 anti-Rabbit IgG is site-specifically conjugated to two fluorophores per sdAb (FluoTag®–X2 variant). We also offer FluoTag®-X4 anti-Rabbit IgG, a blend of two different sdAbs, each carrying two fluorophores, that bind simultaneously to Rabbit IgG for even brighter signal. To learn more about FluoTags®, please visit our Technology section here.

We provide this product conjugated to various fluorophores and offer custom conjugation services with most commercially available fluorophores, provided these are available in a thiol-reactive activated form.

MSDS
Protocols
Reconstitution and Storage
Multiplexing (MX-IF)
Variations:
Conjugation Amount Cat No. RRID
abberior STAR deepRED 500 μL N2402-AbdeepRED-S -
abberior STAR 550 500 μL N2402-Ab550-S AB_3741785
Janelia Fluor 635b 500 μL N2402-JF635B-S AB_3713469
abberior STAR RED 500 μL N2402-AbRED-S AB_3697324
abberior STAR ORANGE 500 μL N2402-AbORANGE-S AB_3697323
abberior STAR GREEN 500 μL N2402-AbGREEN-S AB_3697322
abberior STAR 635P 500 μL N2402-Ab635P-S AB_3076034
abberior STAR 460L 500 μL N2402-Ab460L-S AB_3697321
Atto488 500 μL N2402-At488-S AB_3076037
Alexa Fluor™ 568 500 μL N2402-AF568-S AB_3076035
Atto643 500 μL N2402-At643-S AB_3076038
Alexa Fluor™ 647 500 μL N2402-AF647-S AB_3076036
LI-COR IRDye 680RD 500 μL N2402-Li680-S AB_3076039
LI-COR IRDye 800CW 500 μL N2402-Li800-S AB_3076040
Related Products:

FluoTag®-X2 anti-Mouse IgG1

FluoTag®-X2 anti-Mouse IgG2a/b

FluoTag®-X4 anti-Rabbit IgG

View related products
Clone: 10E10
Host: Alpaca
Produced in: E.coli
Application: IF, WB
Dilution: 1:500, corresponding to 10 nM final concentration of sdAb (20 nM of fluorophore)
Capacity: N/A
Antigen: -
Targets: Rabbit IgG
Specificity: Recognizes native Rabbit IgG. It does not cross-react with Mouse, Guinea Pig, Chicken, Rat, Donkey, Goat, Pig, Horse, Cow or Human IgGs.
Formulation:

The single sdAb clone was lyophilized from PBS pH 7.4, containing 2% BSA (US-Origin). Reconstitute with 500 µL of sterile 50% glycerol (v/v) in deionized water. We recommend including 0.1% sodium azide as a preservative if applicable. When reconstituted with 500 µL, the single-domain antibody concentration is 5 µM. For more details, click the Protocols button above and check Reconstitution and Storage.
kDa: -
Ext Coef: -
Shipping: Ambient temperature
Storage:

Vials containing lyophilized reagent can be stored at 2-8°C for up to 12 months. After reconstitution, store at -80°C for up to 6 months. Working aliquots can be stored at -20°C for up to 4 weeks. For more details, click the Protocols button above and check Reconstitution and Storage.
Protocols:

Relevant protocols can be found under the Protocols button above. For additional information, visit our Resources page.
References:
  1. Saleeb R, O’Shaughnessy J, Ferguson R, Adams CT, Horrocks MH. Single-molecule validation and optimised protocols for the use of secondary nanobodies in multiplexed immunoassays. J Microsc. 2026;301(3):422-436. doi:10.1111/jmi.70059 (IF; Multiplexing; TIRFM; Single-molecule binding assay; SH-SY5Y cells)
  2. Rahman F, Augoustides V, Tyler E, Daugird TA, Arthur C, Legant WR. Mapping the nuclear landscape with multiplexed super-resolution fluorescence microscopy. Nat Commun. 2025;16(1):6042. Published 2025 Jul 1. doi:10.1038/s41467-025-61358-0
  3. Chowdhury R, Mimoso T, Chouaib AA, et al. Microtubules as a versatile reference standard for expansion microscopy. Commun Biol. 2025;8(1):499. Published 2025 Mar 26. doi:10.1038/s42003-025-07967-3 (IF; ExM)
  4. Markusson S, Raasakka A, Schröder M, et al. Nanobodies against the myelin enzyme CNPase as tools for structural and functional studies. J Neurochem. 2024;169(1):1-25. doi: 10.1111/jnc.1627 (IF; STED; mouse teased sciatic nerves)
  5. Mougios N, Opazo F, Rizzoli SO, Reshetniak S. Trafficking proteins show limited differences in mobility across different postsynaptic spines. iScience. 2023;26(2):105971. Published 2023 Jan 13. doi:10.1016/j.isci.2023.105971 (ICC/IF; rat)
  6. Saal KA, Shaib AH, Mougios N, Crzan D, Opazo F, Rizzoli SO. Heat denaturation enables multicolor X10-STED microscopy. Sci Rep. 2023;13(1):5366. Published 2023 Apr 1. doi:10.1038/s41598-023-32524-5 (ICC/IF; rat)
  7. Kilisch M, Gere-Becker M, Wüstefeld L, et al. Simple and Highly Efficient Detection of PSD95 Using a Nanobody and Its Recombinant Heavy-Chain Antibody Derivatives.Int J Mol Sci. 2023;24(8):7294. Published 2023 Apr 14. doi:10.3390/ijms24087294 (IHC; mouse)
  8. Aktalay Hippchen A. Characterization and Optimization of Fluorescent Probes and Labeling Strategies for Nanoscopy Techniques. PhD thesis (2023). Univeristy of Heidelberg (ICC/IF; SMLM; tested species: human, U2OS)
  9. Babai N, von Wittgenstein J, Gierke K, Brandstätter JH, Feigenspan A. The absence of functional bassoon at cone photoreceptor ribbon synapses affects signal transmission at Off cone bipolar cell contacts in mouse retina. Acta Physiol (Oxf). 2021;231(3):e13584. doi:10.1111/apha.13584 (ICC/IF; mouse)
  10. Saal KA, Warth Pérez Arias C, Roser AE, et al. Rho-kinase inhibition by fasudil modulates pre-synaptic vesicle dynamics. J Neurochem. 2021;157(4):1052-1068. doi:10.1111/jnc.15274 (IF, STED; rat primary neurons)
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
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