Application of Antibody-fluorophore Conjugated Probes

Fluorescent antibody refers to conjugate fluorophores to therapeutic antibodies based on well-established chemical method. Chosen from our extensive catalog of therapeutic antibodies and fluorophores, FluoroAb™ products cover a wide range of disease targets and photochemical properties to suit your needs in immunolocalization of antigen and mechanistic study of intermolecular interactions, for diagnostic or imaging purposes.

Introduction of Fluorophores

Fluorophores are a family of chemical compounds that can emit light upon the adsorption of energy from an excitation light source. For a certain fluorophore, there is a specific excitation and emission spectrum. Typically, the wavelengths of light that can cause the maximum excitation and be captured from the maximum emission (Excitation/Emission) are used to refer to a given fluorophore. Most fluorophores are small organic compounds with 20–100 atoms, but there are also exceptions with larger sizes, e.g., natural fluorescent proteins. Fluorophores can be used either alone, together with other fluorescent agents, or as the fluorescent tags in a conjugate, e.g., a fluorophore-labeled antibody. Fluorophores or fluorescent conjugates play important roles in biochemical and biomedical studies and applications. For example, Fluorophores are widely used as fluorescent labels for fluorescence microscopy and near-infrared spectroscopy. With the development of new technologies, certain fluorophores are exploited for live-cell imaging, single molecule tracking, and high-resolution imagery.

Application of Antibody-fluorophore Conjugated Probes

Generally, certain biomolecules, particularly antibodies, can direct fluorophores to a specific site, allowing them to be used as fluorescent probes for a given target. The large size of an antibody allows multiple fluorophores to be attached on a single antibody, which breaks the intensity limitation of a single fluorophore. The conjugation between fluorophores and antibodies can be achieved via specific interactions between functional groups on both components, e.g., amino-succinimide reaction, thiol-maleimide reaction, as well as click chemistry.

Flow cytometry is a popular application of antibody-fluorophore conjugated probes. Fluorophore labeled antibodies enable cell sorting techniques such as fluorescence-activated cell sorting (FACS) flow cytometry for the fast and efficient identification and collection of cells expressing the desired surface antigen. In the meantime, with the development of more advanced imaging flow cytometry platforms, antibody-fluorophore probes not only facilitate the sorting and quantification of surface antigen, but also provide the means for the acquisition of high-resolution images of the cells bearing the antibody binding targets.

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Advantages of FluoroAb™ Products

Compared to indirect methods such as a secondary fluorescently labeled antibody, attaching fluorescent dyes directly to the antibody offers unique advantages:

  • Quicker and more reproducible results
  • Suitable for multi-protein complexes
  • No need for secondary antibodies
  • No disturbance to epitope recognition
  • Enhanced robustness
  • Low batch-to-batch variation

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What Do We Provide?

At Creative Biolabs, we provide all kinds of FluoroAb™ products conjugated by our abundant therapeutic antibodies and fluorophores. Here, a large variety of organic fluorophores covering a broad excitation spectrum from visible to near-infrared range are available, including Alexa Fluor 350, 405, 488, 532, 546, 555, 568, 594, 647, 680, 750, and Bodipy FL, Coumarin, Cy3, Cy5, Fluoresein, Oregon green, Pafific blue…Besides, with expertise in antibody development, Creative Biolabs provides customers with a broad range of antibodies for selection.

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