Immuno-PCR Assays
Achieve higher sensitivity than is possible using a standard ELISA with RayBiotech's Immuno-Quantitative PCR ELISA (IQELISA) and Bead-based Immuno-Quantitative PCR ELISA (BIQ-ELISA).
What is an Immuno-PCR Assay?
The IQELISA and BIQ-ELISA are immuno-PCR assays that combine the specificity and ease of use of an ELISA with the sensitivity of real-time PCR (RT-PCR, also referred to as quantitative PCR or qPCR), providing you with a detection platform that uses familiar ELISA workflows while pushing the frontiers of your detection limits and sample volume requirements.
The benefits of an immuno-PCR kit include over a standard ELISA include:
- Up to 1000x more sensitivity—As with all ELISAs, assay sensitivity depends on the sensitivities of the antibodies being used. However, the lowest limits of quantification (LLOQs) of IQELISA are, on average, 23-fold lower than standard ELISA with the same antigen-antibody pair, and can get up to 1000-fold lower for BIQ-ELISA.
- Sample volume requirements as low as 10 µL—Compared to a standard ELISA, immuno-PCR kits require 1/10th the sample volume, ensuring quantitative detection even when sample is limiting.
How Does Immuno-PCR Work?
IQELISA
- RayBio® IQELISA 96-well PCR plates are pre-coated with specific capture antibodies.
- To start, pipette standards and samples into the wells. The target protein in the standards and the samples will bind to the immobilized antibody.
- Wash the wells and add the detection affinity reagent which will bind to any captured antigen.
- Finally, place the plate into a qPCR instrument for cycling and measurement of DNA amplification. The cycle number where amplification is detected is proportional to the amount of affinity detection reagent bound to captured antigen in each well.
BIQ-ELISA
- RayBio® BIQ-ELISA contain beads pre-coated with specific detection antibodies.
- To start, mix the antibody-conjugated beads with oligo-conjugated antibody and pipette into wells.
- Pipette the standards and samples into the wells. The target protein in the standards and samples will bind to both the bead-immobilized antibody and the oligo-conjugated antibody to form a sandwich complex.
- Wash the wells and add elution buffer to elute the complex and transfer to a 96 well PCR plate.
- Finally, place the plate into a qPCR instrument for cycling and measurement of DNA amplification. The cycle number where amplification is detected is proportional to the amount of oligo-conjugated antibody bound to antigen in each well.
Save Time With Our ELISA Services
If you’d like to get the data from an IQELISA study but are resource limited, you can take advantage of the experienced scientists who run our ELISA Services studies. Simply send us your samples and we will perform full testing using our IQELISA immuno-PCR kits in our GLP-compliant laboratory. Once it’s complete, we’ll send you a full report.
If you need even more sensitivity, try our ultrasensitive biomarker testing service using Simoa® technology. Like IQELISA, Simoa® is orders of magnitude more sensitive than standard sandwich-based immunoassay techniques. RayBiotech's Ultrasensitive Simoa Testing Services can achieve sensitivity as low as femtogram (fg/mL) levels, allowing you to detect and quantify biomarkers at concentrations previously difficult or impossible to measure.
Learn about all of our ELISA Services >
