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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 sdAb (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 Abberior STAR 580 anti-rabbit IgG sdAb (Cat. No. N2402-Ab580-S). Nuclei were visualized by DAPI staining (blue).

FluoTag®-X2 anti-Rabbit IgG

Cat No: N2402 Category:

195,00 €

This new FluoTag®-X2 anti-Rabbit IgG is a species-specific FluoTag® directed against IgGs from Rabbit. Especially useful for One-step immunofluorescence (IF) and Multiplexing applications.

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Like many other mammals, the immune repertoire of rabbits consists of several immunoglobulin (Ig) isotypes (IgA, IgG, IgE, and IgM). However, compared to other species, rabbits possess only one type of IgG carrying either a lambda or kappa light chain. Historically, rabbit antibodies have been a rich source of polyclonal antibodies, which were employed in various fields of life sciences. They are used either directly from animal sera, or as affinity-purified reagents.  In addition, advances in antibody engineering over the last decade enabled the generation of monoclonal rabbit antibodies. Independent of its clonality and irrespectively of its VH domain, our alpaca-derived single domain antibody (sdAb, or nanobody) binds the constant domain (Fc) of any rabbit IgG, specifically and strongly, which makes this secondary tool ideal for demanding imaging applications.

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 500 μl N2402-At488-S AB_3076037
AZDye568 500 μl N2402-AF568-S AB_3076035
Atto643 500 μl N2402-At643-S AB_3076038
Alexa647 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
AbberiorStar635P 500 μl N2402-Ab635P-S AB_3076034
Related Products:

FluoTag®-X2 anti-Mouse IgG1

FluoTag®-X2 anti-Mouse IgG2a/b

FluoTag®-X4 anti-Rabbit IgG
Clone: 10E10
Host: Alpaca
Produced in: E.coli
Application: IF, WB
Dilution: 1:500 (corresponding to 10 nM final concentration if the sdAb was reconstituted as proposed bellow)
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 50 % glycerol 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
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:

Suggested general protocols for immunofluorescence can be found in our Resource Section.

One-Step immunofluorescence and multiplex staining can be performed with our smart secondaries. Here you can find more on these technologies One-Step-IF & Multiplexing.
References:
  1. 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)
  2. 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)
  3. 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)
  4. 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)
  5. 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.
Legal terms: By purchasing this product you agree to our general terms and conditions.

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