Suggested Application

Multiplexed Protein Detection; Biomarker Screening; Identifying Key Factors; Confirming Biological Process; Biomarker Validation; Validation of Antibody Array Results; Quantitative Protein Detection; Establishing Normal Range

Kit Components

• RayPlex Multiplex Bead Cocktail
• 1X Assay Diluent
• 20X Wash Buffer
• RayBio^® Serum/Plasma Diluent
• Lyophilized Protein Standard Mix
• 1X Biotinylated Detection Antibody Cocktail
• 10X PE Conjugate
• Flow Cytometer Setup Bead Cocktail
• V-bottom 96-well Microplate
• Filter 96-well Microplate

Other Materials Required

• Orbital 96-well plate shaker (with ability to reach 1000 rpm)
• Flow cytometer with blue and red lasers capable of measuring phycoerythrin (PE) and allophycocyanin (APC) channels
• Aluminum foil or 96-well Plate Aluminum Sealers
• Distilled water
• 1.5 ml polypropylene microcentrifuge tubes or similar
• Microcentrifuge
• Recommended: Vacuum manifold compatible with a 96-well microplate if using the Filter 96-Well Microplate
• Optional: 96-well round bottom plate (to prepare samples)
• Optional: High throughput sampler (HTS) for 96-well plate reading

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Protocol Outline

1. Prepare Reagents, Standards, and Samples
2. Add RayPlex Multiplex Bead Cocktail to each well
3. Incubate with Samples and Standards
4. Incubate with Biotinylated Detection Antibody Cocktail
5. Incubate with Streptavidin-Conjugated Fluorophore
6. Analyze with a compatible flow cytometer
7. Perform data analysis

Storage/Stability

Upon receipt, the Lyophilized Protein Standard should be stored at -20°C or below. Other components should be stored at 4°C. Use reagents immediately after preparation. Additional vials of the Lyophilized Protein Standard Mix (Item 5) can be purchased separately. If stored at the proper temperature, the kit will retain complete activity for at least 6 months.