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Ultrasensitive Immunofluorescence Assay

BcMag™ TR-FRET Assay is a time-resolved fluorescence resonance energy transfer assay that uses time-resolved fluorescence magnetic beads as the donor fluorophore. It allows for the detection of trace amounts of analytes in a sample, resulting in higher sensitivity, and eliminates background fluorescence caused by the sample and plastic microplate. This assay offers advantages in high throughput screening, such as assay flexibility, dependability, increased assay sensitivity, higher throughput, and fewer false positive/false negative results. It does not require washing steps.

How It Works

Mix the antibody-conjugated donor beads with the cell lysates and incubate them with continuous rotation for a sufficient time. The beads remain suspended in the sample solution during mixing, allowing the target analytes to bind to the donor beads.
After incubation, the beads are collected and separated from the sample using a magnet rack…
Add the antibody-conjugated acceptor and incubate them with continuous rotation for a sufficient time.
Analysis of TR-FRET measurements.

Advantages and Benefits

1. The ability to perform separation / preconcentration and detection of analytes simultaneously on the same beads, resulting in quick, simple, robust, and high-throughput analysis of trace amounts from complex biological samples.

2. Ultra-sensitive detection limits of 10 pg/mL compared to typical fluorometric detection limits of 100 pg/mL. Extremely photostable and highly resistant to photobleaching.

3. Very high fluorescence intensity due to the high concentration of lanthanide chelate with a high quantum yield.

4. Lanthanide chelate or cryptate has large Stokes shifts, narrow emission bands, and long fluorescence lifetime, which reduces background and increases the signal-to-noise ratio.

5. Many bioprocess ELISA assays can be converted to an HTRF assay.

6. The assay does not require washing steps.

7. The beads have a hydrophilic surface that can be grafted with different functional groups and linkers, allowing for efficient conjugation of various ligands.

8. The magnetic properties of the beads make them suitable for high-throughput automation.