Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2
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Fully defined & ready-to-go fluorescent camelid single-domain antibody (sdAb) conjugates featuring FluoTag® and Smart Secondaries® technologies.
- Quantitative and reliable readout in all fluorescence-based analyses
- Defined number of fluorophores enhance brightness of signals
- Improved localization accuracy ideal for super resolution imaging
- Recognizes conventional immunoglobulins with exquisite species and isotype specificity
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2 (AB325308)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized, PBS washing, and blocked with 10% normal goat serum in PBS HeLa (human cervical cancer cells) cells labelling Actin with ab213251 at 1 : 400 (1.20 µg/ml) dilution, followed by ab325308 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2 at 1/500 (0.15 µg/ml per sdAb) dilution (magenta).
Confocal image showing cytoplasmic staining in HeLa cells (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a fluorescence microscope (Zeiss Axio Imager Z1).
Secondary antibody only control : Secondary antibody is ab325308 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2 at 1/500 (0.15 µg/ml per sdAb) dilution.
- Other
Supplier Data
Other - Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2 (AB325308)
Smart Secondary®
Smart Secondary® sdAb generally recognize the Fc domain on the respective primary antibody (the only exception being the FluoTag-X2 anti-Mouse Kappa light chain). Due to the inherent symmetry of the target antibody, two secondary nanobodies will be able to bind each primary antibody. Our secondary FluoTag®-X2 reagents will therefore target 4 fluorophores to each primary antibody. Our anti-Rabbit IgG tools are also available as FluoTag®-X4.
This reagent is a blend of two sdAbs independently binding the same target molecule. As each target site is present twice and each sdAb carries two fluorophores, this reagent can target up to eight fluorophores to each primary antibody – ideal for bright images!
Reactivity data
Product details
FluoTags®
FluoTags are fully defined & ready-to-go fluorescent camelid single-domain antibody (sdAb) conjugates. All FluoTags® carry a defined number of fluorophores at known positions. Accordingly, they are available as FluoTag®-X2 or FluoTag®X4.
Together with their monovalent binding mode, the defined fluorophore conjugation of FluoTags® results in a more quantitative and reliable readout in all fluorescence-based analyses. Due to the small size the fluorophore displacement from the bound target protein is very low and therefore FluoTags® are ideal for standard super resolution technologies.
FluoTag®-X2 (One target - One sdAb – Two fluorophores (1:1:2)
FluoTag®-X2 reagents are composed of a single sdAb carrying two fluorophores per protein molecule. The maximal displacement of fluorophores from the antigen-binding surface is ~3 nm. FluoTag®-X2 reagents show good tissue penetration features.
Features:
- High affinity monovalent binding + two fluorophores per sdAb for brighter signals
- Reagent is made from small-sized proteins. This ensures optimal access to the respective target protein, also in crowded environment or 3D samples.
Smart Secondaries®
Smart Secondaries® are a unique set of high-affinity single-domain antibodies recognizing conventional immunoglobulins with exquisite species and isotype specificity. They provide unique benefits for all antibody-based assays in research and diagnostics by combining reproducible specificity and production with small and high-affinity monovalent binding. The technology offers surprising new possibilities like one-step indirect immunofluorescence or easy multiplexing.
In comparison to conventional secondary reagents, Smart Secondary® nanobodies feature significant advantages:
- Small probes for enhanced staining speed and improved tissue penetration and higher resolution.
- Monoclonal tools: Clearly defined species specificity – once and forever!
- Isotype specificity for independent detection of mouse IgG1 and IgG2a/b isotypes.
- Recombinant production for reliable quality, scalability and reproducibility without using animals.
- Monovalent binding mode – Pre-incubation without clustering primary antibodies and therefore one-step indirect immunofluorescence possible!
- Extraordinarily strong binding allows one-step multi-colour staining and imaging even with primary antibodies from identical species and isotype (Multiplexing)
- Equipped with propriety FX technology for improved post-fixation and enhanced performance in demanding application (e.g., expansion microscopy, DNA-PAINT, dSTORM, etc.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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