Product Ordering Information
Substrate selection is a key component in developing ELISAs with high signal and low background. LI-COR offers proprietary HRP 680 and AP 680 Substrates that are optimized for use in the near-infrared region (680 nm) on the Aerius, and Odyssey® CLx, Odyssey Classic, and Odyssey Sa Infrared Imaging Systems.
Near-infrared (NIR) detection overcomes the limitations of chromogenic substrate detection, which does not allow for quantification of greater than four optical density units. NIR detection allows for a wider linear range.
LI-COR ELISA HRP Substrate (680) and ELISA AP Substrate (680) offer equal or better sensitivity compared to commercially-available chromogenic substrates and are ideal for endpoint assays. The substrates provide excellent signal-to-noise ratios, consistency, and linearity, making them ideal for use in most ELISA applications.
Figure 1. Serial dilutions of rabbit IgG detected with Goat Anti-Rabbit-HRP followed by LI-COR ELISA HRP Substrate (680) (P/N 926-34300). Row 1 (blank) was coated with PBS only. Rows 2-5 were coated with serial dilutions (5000 - 0.0006 ng/mL) of rabbit IgG in PBS (rows 2 and 3 are duplicates and rows 4 and 5 are duplicates). Imaged on the Odyssey Infrared Imager.
Many commercially-available microwell plates and strip wells designed for ELISA or EIA/RIA use are compatible with the Aerius, and Odyssey family of imaging systems. For optimal ELISA performance with near-infrared fluorescent detection, clear, flat-bottomed plates are required. Use of flat-bottomed plates that are not clear will result in signal not being detected. It is advisable to scan the plate prior to addition of substrate to ensure the plate does not fluoresce in the 700 nm channel.
NOTE: These products do not work in Western blotting applications.