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AB325308

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
2 Images
Immunocytochemistry/ Immunofluorescence - Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2 (AB325308)
  • 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 - Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) - FluoTag®-X2 (AB325308)
  • 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!

Key facts

Host species

Alpaca

Target species

Human

Target isotype

IgG1, IgG4

Target specificity

Fc region

Minimal cross-reactivity
Pre-adsorbed

No

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Applications

ICC/IF

applications

Clonality

Multiclonal

Clone number

2F3 / 2E5

Isotype

IgG

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ICC/IF": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"1/500", "notes":"<p></p>" } } }

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:

  1. Small probes for enhanced staining speed and improved tissue penetration and higher resolution.
  2. Monoclonal tools: Clearly defined species specificity – once and forever!
  3. Isotype specificity for independent detection of mouse IgG1 and IgG2a/b isotypes.
  4. Recombinant production for reliable quality, scalability and reproducibility without using animals.
  5. Monovalent binding mode – Pre-incubation without clustering primary antibodies and therefore one-step indirect immunofluorescence possible!
  6. Extraordinarily strong binding allows one-step multi-colour staining and imaging even with primary antibodies from identical species and isotype (Multiplexing)
  7. 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.

Properties and storage information

Form
Liquid
Purification technique
Size-exclusion chromatography
Storage buffer
Constituents: PBS, 50% Glycerol (glycerin, glycerine)
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1 month
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-80°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Product protocols

Target data

Immunoglobulins, also known as antibodies, are membrane-bound or secreted glycoproteins produced by B lymphocytes. In the recognition phase of humoral immunity, the membrane-bound immunoglobulins serve as receptors which, upon binding of a specific antigen, trigger the clonal expansion and differentiation of B lymphocytes into immunoglobulins-secreting plasma cells. Secreted immunoglobulins mediate the effector phase of humoral immunity, which results in the elimination of bound antigens (PubMed:20176268, PubMed:22158414). The antigen binding site is formed by the variable domain of one heavy chain, together with that of its associated light chain. Thus, each immunoglobulin has two antigen binding sites with remarkable affinity for a particular antigen. The variable domains are assembled by a process called V-(D)-J rearrangement and can then be subjected to somatic hypermutations which, after exposure to antigen and selection, allow affinity maturation for a particular antigen (PubMed:17576170, PubMed:20176268).
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