Product #: R001

High Intensity Chemiluminescence ELISA Substrate

HaemoScan High Intensity HRP-Chemiluminescence ELISA Substrate uses an enhanced chemiluminescence formula which results in luminescence signals up to four times the intensity of the top competitor.

European Union Price (As per 01-01-2016):
100 mL: €80,-
250 mL: €190,-
500 mL: €360,-
1000 mL (2x 500 mL): €495,-
Inquire for a quotation if you are located outside the European Union.


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Chemiluminescence Substrate
Description

HaemoScan High Intensity HRP-Chemiluminescence ELISA Substrate uses an enhanced chemiluminescence formula which results in luminescence signals up to four times the intensity of the top competitor and up to 13 times the intensity of a standard Luminol + Phenol based substrate [1] (Figure 1), providing high sensitivity during ELISA. The luminescence signal of this substrate reaches its maximum within minutes (Figure 2). Luminescence based ELISAs are usually performed in black or white opaque microplates, but may also be performed in tubes.

Chemiluminescence Substrate

Figure 1. High intensity light signal with the luminol-based HaemoScan High Intensity HRP-Chemiluminescent ELISA Substrate. Intestinal fatty acid-binding protein (IFABP) (aka: Fatty acid-binding protein 2 [FABP2]) calibrators were analyzed at room temperature by means of enzyme-linked immunosorbent assay (ELISA) using a white opaque Nunc Maxisorp microplate. After the final incubation step (Streptavidin-HRP), the microplate was washed with Phosphate-buffered Saline + 0.10% Tween 20, pH 7.4 and 100 µL substrate was added per well. After incubation on a plate shaker for 10 seconds, luminescence was measured with a gain 110 after one minute (GENios, Tecan Trading AG, Switzerland).

Chemiluminescence Substrate

Figure 2. Rapid light generation with the luminol-based HaemoScan High Intensity HRP-Chemiluminescence ELISA Substrate. Streptavidin-HRP was diluted to 800 pg/mL in 0.1M Tris, pH 8.5 and 50 µL was transferred to a white opaque Nunc Maxisorp microplate. After addiction of 50 µL HaemoScan High intensity HRP-chemiluminecence ELISA substrate, the microplate was incubated at room temperature on a shaker for 10 seconds and luminescence was measured with a gain of 120 at one minute intervals (GENios, Tecan Trading AG, Switzerland).

Features
  • High signal intensity (up to 13x signal intensity of a standard Luminol + Phenol based substrate [1])
  • Rapid light generation (maximum signal is produced within minutes)
  • Large dynamic range (detection down to picogram levels)
  • High signal:noise ratio (low background)
  • Convenient handling (easy-to-use 1:1 mixture of two components)
  • Convenient storage (ambient shipping and storage conditions)
Application

HaemoScan High Intensity HRP-Chemiluminescence ELISA Substrate is an enhanced chemiluminescence substrate for the detection of horseradish peroxidase (HRP) activity, optimized to detect picogram level proteins in ELISA.

Principle

Peroxidases such as horseradish peroxidase catalyze the oxidation of luminol to 3-aminophthalate in the presence of a catalyst such as perborate. This reaction is accompanied by the emission of light at 425 nm. Chemiluminescent based ELISA’s are quantified by using a luminometer to measure the relative light units (RLU). In contrast to colorimetric (chromogenic) substrates which produce a colored product that persists after the enzyme-substrate reaction has occured, chemiluminescence substrates only produce light during the enzyme-substrate reaction and can be measured immediately.

Storage and Stability

Product is stable at room temperature for at least one year.

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Manual / SDS

References

[1] G. H. Thorpe, L. J. Kricka, S. B. Moseley, and T. P. Whitehead, “Phenols as enhancers of the chemiluminescent horseradish peroxidase-luminol-hydrogen peroxide reaction: application in luminescence-monitored enzyme immunoassays.,” Clin. Chem., vol. 31, no. 8, pp. 1335–41, Aug. 1985.