Luminata™ ELISA HRP Substrates
Sensitivity and dynamic detection range for even your most demanding assays
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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 |
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Luminata ELISA HRP Substrates are more sensitive than TMB substrates
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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.