Luminata™ ELISA HRP Substrates
Recommended Products
Overview
The Luminata ELISA HRP chemiluminescent substrates provide three major advantages:
- High sensitivity for detecting as little as 0.1 pg of protein
- Broad dynamic detection range (up to 2,500-fold), enabling a wide range of quantitation.
- Flexibility to adjust ELISA signal strength by two methods:
a. Exposing the well to the luminometer for a longer time
b. Applying a more sensitive ELISA substrate to obtain signals in wells with otherwise undetectable signals.
Key Features of Luminata ELISA HRP substrates
| Luminata Crescendo ELISA |
Luminata Forte ELISA |
Tetramethylbenzidine (TMB) | |
| Limit of detection* | ~0.4 pg | ~0.1 pg | > 2.4 pg |
| Dynamic detection range | ~625x (0.4-250pg) |
~2500x (0.1-250pg) |
~260x (2.4 -625pg) |
| Signal half-life** | >1 hr | >1 hr | Permanent |
| Stability of working solution (after mixing) | >8 hrs | >8 hrs | N/A |
| Can re-apply a different substrate | Yes | Yes | No |
| Shelf life | 1 year | 1 year | 6 months |
| Incubation time | 1 – 5 min | 1 – 5 min | >15 min |
| * Based on IL-2 sandwich assay using 10 μg/ml capture antibody and 0.5 μg/ml detection antibody. ** Based on IL-2 sandwich assay using ~16 pg of protein/well. N/A = Not Applicable |
|||
Luminata ELISA HRP Substrates are more sensitive than TMB substrates
Luminata Crescendo and Forte ELISA HRP substrates are more sensitive than TMB substrate. Detection limits for each ELISA substrate were measured with sandwich IL-2 ELISA titration assays using an IL-2 ELISPOT antibody pair (Millipore Cat. No. ELI-002-M). Luminescence was measured using a Wallac VICTOR2™ 1420 counter and TMB color was measured at absorbance 450 nm.
Luminata ELISA HRP Substrates are the most sensitive in their class
Luminata ELISA HRP substrates offer higher sensitivity than other chemiluminescent ELISA HRP substrates. Three HRP chemiluminescent substrates for ELISA were tested in sandwich IL-2 (A) or indirect GAPDH (B) ELISA assays. Purified GAPDH (Sigma G5262-1VL) was assayed using mouse anti-GAPDH (Millipore Cat. No. MAB374) and goat anti-mouse IgG, HRP (Millipore Cat. No. AP124P). Luminescence was measured using a Wallac VICTOR2™ 1420 counter.