DyLight-conjugated anti-mouse primary antibodies are directly labeled immunoreagents that combine a mouse antigen-specific immunoglobulin with a DyLight fluorescent dye. DyLight Fluors are a complete family of high-intensity, photostable fluorescent tags developed for labeling antibodies and other molecular probes. The dyes have absorption maxima ranging from 350 nm to 777 nm, covering the entire visible light spectrum and several key near-infrared and infrared wavelengths.
The key advantages of DyLight dyes over conventional fluorophores include higher fluorescence intensity and photostability than Alexa Fluor, CyDye, and LI-COR dyes in many applications. DyLight dyes remain highly fluorescent over a broad pH range (pH 4–9), and their water solubility allows a high dye-to-protein ratio to be achieved without causing precipitation of conjugates. These properties make DyLight conjugates particularly suitable for applications requiring prolonged illumination or demanding pH conditions.
Applications
Flow Cytometry
DyLight-conjugated anti-mouse primary antibodies are validated for flow cytometry. The anti-mouse CD4 clone GK1.5 conjugated to DyLight 488 is specifically validated for flow cytometry, immunohistochemistry, and immunocytochemistry, with applications including T helper cell and regulatory T cell analysis. The excitation laser for DyLight 488 is the blue laser at 493 nm. For flow cytometric staining, the suggested use of DyLight-conjugated reagents is typically ≤1.0 µg per 10⁶ cells in 100 µL volume.
Immunofluorescence and Immunocytochemistry
DyLight-conjugated anti-mouse antibodies are widely used for immunofluorescence microscopy. VectaFluor Duet Immunofluorescence Double Labeling Kits utilize DyLight 488 anti-mouse (green) and DyLight 594 anti-rabbit (red) in a single-step detection reagent for simultaneous detection of mouse and rabbit primary antibodies. The DyLight 594 goat anti-mouse IgG conjugate delivers noticeably brighter fluorescence than Alexa Fluor 594 and much brighter, more water-soluble performance than Texas Red, with validated applications in flow cytometry, immunocytochemistry, and immunofluorescence at recommended dilutions of 1:50–1:1,000.
Western Blotting and ELISA
DyLight-conjugated anti-mouse secondary antibodies are also employed in Western blotting and ELISA, with working dilutions typically ranging from 1:500 to 1:20,000 depending on the product and application. The high water solubility of DyLight dyes reduces aggregation and non-specific binding, contributing to low background in these assays.
