Introduction
RayPlex® is a multiplexed, sandwich- and bead-based quantitative antibody array for the simultaneous detection of multiple proteins using small sample volumes. It couples RayBiotech's antibody pair library with reliable flow cytometry, requiring no dedicated instrumentation—only a common flow cytometer equipped with blue (or green) and red lasers. Capture antibodies are immobilized on beads with distinct fluorochrome/intensity combinations; target proteins are detected via a biotinylated detection antibody and PE conjugate, and quantified against a standard curve of purified protein standards. Minimal sample volume is 25 µL per test after dilution.
Product Features
- Less sample, more data: up to 13 proteins quantified from only 25 µL or less of your precious sample
- ~4 hour processing time
- More cost-effective than running individual ELISAs
- Compatible with most flow cytometers; can also be used with Luminex® systems
- No specialized or dedicated instrument(s) needed
- High specificity (no detectable cross-reactivity between antibodies or targets) and system reproducibility
- Well-suited for high-throughput assays
Targets Detected
PD1 (PDCD1), BTLA (CD272), VISTA (B7-H5 / PD-1H / VSIR), CTLA-4 (CD152 / CTLA4), CD28, ICOS, ILT-4 (LIR-2 / CD85d / LILRB2), TIM-3 (KIM-3 / HAVCR2), TIGIT, 4-1BB (TNFRSF9 / CD137), GITR (TNFRSF18), ILT-2 (LIR-1 / CD85j / LILRB1), TIM-1 (KIM-1 / HAVCR1)
Kit Components
- RayPlex® Magnetic Multiplex Bead Cocktail
- Assay Diluent F
- Wash Buffer (20X)
- Human Serum/Plasma Diluent
- Lyophilized Protein Standard Mix
- Biotinylated Detection Antibody Cocktail
- PE Conjugate (10X)
- Flow Cytometer Setup Bead Cocktail
- V-Bottom 96-well Microplate
- Flat-Bottom 96-well Microplate
Additional Materials Required
- Orbital 96-well plate shaker (capable of 1000 rpm)
- Flow cytometer with blue (or green) and red lasers capable of measuring PE, Alexa Fluor 700 (AF700), and Alexa Fluor 647 (AF647)/APC channels, OR a Luminex instrument
- Aluminum foil or 96-well plate aluminum sealers
- Deionized (DI) water
- 1.5 mL polypropylene microcentrifuge tubes or similar
- Microcentrifuge
- Recommended: Automated plate washing system with magnetic separation
- Optional: 96-well round-bottom plate, high throughput sampler (HTS), or vacuum manifold for a filter microplate
Assay Procedure Summary
- Prepare reagents, standards, and samples
- Add the RayPlex® Magnetic Multiplex Bead Cocktail to each well
- Incubate with samples and standards (2 hours at room temperature, or overnight at 4°C)
- Wash, then incubate with the Biotinylated Detection Antibody Cocktail (1 hour)
- Wash, then incubate with PE Conjugate (30 minutes)
- Wash, resuspend, and analyze on a compatible flow cytometer or Luminex instrument
Storage/Stability
Upon receipt, store the Lyophilized Protein Standard Mix at -20°C or below; store all other components at 4°C. Use reagents immediately after preparation. If stored at the proper temperature, the kit will retain complete activity for at least 6 months.