Step-by-step guide to integrating Exazym’s BOLD amplification into your existing ELISA workflows
We’re excited to announce the release of a comprehensive protocol for implementing ELISA signal amplification in standard 96-well plate applications. This step-by-step guide provides researchers with detailed guidance for integrating Exazym’s BOLD (Binding Oligo Ladder Detection) signal amplification into traditional ELISA workflows.
One of Exazym’s key strengths is compatibility with existing lab infrastructure. Many researchers already have ELISA workflows optimized for their specific biomarkers. Our new protocol makes it straightforward to enhance those assays with BOLD amplification—without starting from scratch.
Why This Matters
When you implement signal amplification in your existing ELISA assays, you’re not reinventing your entire workflow. Instead, you’re layering in proven amplification chemistry that works within your established systems.
Researchers often face a difficult choice: redesign everything to achieve better sensitivity, or accept the limitations of their current workflows. With our ELISA signal amplification protocol, you eliminate that tradeoff. By following our step-by-step guidance, you can integrate BOLD technology into existing ELISAs while preserving your validated antibody pairs, buffer systems, and equipment.
The result is meaningful sensitivity improvement—often 10-fold to 100-fold gains—without operational friction or equipment replacement. For more context on why signal amplification matters in modern immunoassay development, read our white paper on BOLD amplification in immunoassays.
Key Features of the ELISA Signal Amplification Protocol
Two Implementation Paths
The protocol guides you through two distinct approaches:
- Adapting existing ELISAs: Use the same coating, blocking, and incubation procedures as your current standard ELISA, then layer in Exazym’s amplification steps. This is the fastest path to improved sensitivity if your assay is already running.
- Developing new assays: Starting from the ground up? The protocol provides recommended concentrations and conditions for capture antibodies, detection antibodies, and washing procedures optimized for amplification.
Practical Guidance Includes
- Detailed antibody concentration recommendations (0.25-6 µg/mL for capture, 0.025-0.5 µg/mL for detection)
- Buffer and blocking solutions that work well with Exazym
- Step-by-step polymerization procedure (just 30 minutes at room temperature)
- Troubleshooting tips for reducing background and optimizing sensitivity
- Complete reagent formulations ready to implement
For detailed information on optimizing your specific biomarker assay, explore our collection of application notes demonstrating signal amplification across different targets.
What You Need to Get Started
The protocol works with straightforward, commonly available components:
- Standard Maxisorp ELISA plates (96-well or strips)
- Your existing ELISA reader
- Exazym Detection Kits (the protocol references specific components)
- Common laboratory reagents
No specialized instrumentation. No software licensing. Just the protocol, your existing equipment, and Exazym’s BOLD amplification chemistry.
The Real Benefit
You don’t need to redesign your assay or invest in new equipment. If you’re already running ELISAs, you can integrate amplification in about 30 additional minutes—achieving the sensitivity improvements you need while preserving your validated workflow.
This is the practical reality of working with BOLD: enhanced sensitivity without operational disruption.
Getting Started
Ready to enhance your ELISA sensitivity? Access the BOLD ELISA Protocol, or if you’d like guidance implementing it for your specific biomarkers, we’re here to help.
Reach out to us—these discussions about your actual assays are where we can make the biggest difference.