One-Step Dye Removal Magnetic Beads

Introduction

Fluorescent dyes, also called reactive dyes or fluorophores, are natural or synthetic compounds that absorb light and re-emit it at a longer wavelength. Due to their unique advantages such as versatility, sensitivity, and quantitative capabilities, fluorescent dyes are widely used to label biologically relevant molecules such as protein, antibodies, peptides, DNA, and RNA for their applications in cell biology, immunology, biochemistry, microbiology, molecular biology, genomics, and proteomics. After a fluorescent labeling reaction, removing excess or unreacted dye from final conjugates is often necessary since it interferes with many downstream applications. Removing fluorescent dyes is usually accomplished by spin columns, gel filtration, gravity-flow columns, and dialysis. However, those traditional methods present many problems, including time-consuming and labor-intensive processes, poor recovery of protein, peptides, or nucleic acids, and the challenge of adapting to automation. For this reason, we introduce a novel one-minute dye removal system.

Hygia One-Step Dye Removal Kit uses specially formulated resin with a proprietary surface chemistry to specifically remove the excess free (non-conjugated) fluorescent dyes from the finished protein, peptide or antibody labeling reaction. Compared with the dye removal columns, the resin can quickly and efficiently remove free dyes from the sample with just a single step and enables an individual or 96 samples to be processed simultaneously in less than 1 or 10 minutes with very little hands-on time. Since the magnetic resin only adsorbs the free dye, the labeled biomolecule recovery rate is exceptionally higher than >90%. Moreover, the magnetic beads can remove most of the dyes if the appropriate amount of samples and buffer conditions are used.

Workflow

The one-minute dye removal protocol is straightforward.

1. Add the beads directly to the sample.

2. Pipette or vortex to capture the free dye.

3. Magnetic separation of the beads from the protein solution, while the supernatant contains the purified and ready-to-run products.

Features and Advantages

● Simple protocol: No liquid transfer, One-tube, One-step, and One-minute protocol

● Easy-to-use

● Reliable and reproducible results with exceptional >90% recovery for protein (>6 kDa, aprotinin) or DNA/RNA (>25mer dsDNA)

● Effective Cleanup: Remove 95% free dye

● Cost-effective: Eliminates columns, filters, and laborious repeat pipetting

● High-throughput: Compatible with many different automated liquid handling systems