DyLight-conjugated anti-human primary antibodies are fluorescent immunoreagents utilizing a family of high-performance synthetic dyes for detecting human antigens across multiple spectral ranges. Produced by immunizing hosts such as goat, rabbit, or mouse with purified human immunoglobulins or specific human proteins, these antibodies are affinity-purified and covalently conjugated to DyLight fluorophores via NHS-ester chemistry targeting primary amines. The DyLight series spans the visible to near-infrared spectrum, including DyLight 488 (green, ex/em 493/518 nm), DyLight 550 (yellow, ex/em 562/576 nm), DyLight 594 (red-orange, ex/em 593/618 nm), DyLight 633 (red, ex/em 638/658 nm), DyLight 650 (far-red, ex/em 652/672 nm), and DyLight 800 (infrared, ex/em 777/794 nm).
These dyes exhibit high fluorescence intensity and photostability compared with commonly used fluorophore conjugates such as Alexa Fluor, CyDye, and FITC in many applications, while maintaining fluorescence across a broad pH range of 4–9. High water solubility enables high dye-to-protein ratios while minimizing precipitation. Available specificities include anti-human IgG (H+L), anti-human IgG (Fc), isotype-specific reagents, and anti-human CD markers. Extensive cross-adsorption helps minimize nonspecific binding.
Applications
DyLight conjugates are employed across diverse fluorescence-based platforms. In immunofluorescence microscopy and confocal imaging, their photostability supports prolonged imaging and multicolor experiments, while defined emission spectra can help minimize spectral bleed-through. In flow cytometry and fluorescence-activated cell sorting (FACS), high fluorescence intensity facilitates the detection of low-abundance human antigens in multicolor panels. Additional applications include immunohistochemistry, ELISA, high-content screening, and Western blotting.
Working dilutions typically range from 1:100 to 1:2,000 depending on the application. Store at 4°C, protected from light; do not freeze. Intended for research use only.
