Elisa

The smallest detectable dose in the ELISA is below 0.1pg/well. A typical standard curve is shown in Figure 2. The intra-assay variation for the assay in cell culture medium (without extraction) is between 4 and 9%. In cell culture media and C18-extracted samples the recovery is usually 100%, provided that standards and samples contain a similar matrix and received the same treatment (data not shown).

Table 1. Cross-reactions of various indole derivatives measured in the RIA system

Indole derivative

Cross-reaction (%)

Melatonin

100.0

6-Hydroxy-melatonin

0.800

5-Methoxy-tryptamine

0.030

N-Acetyl-5-hydroxy-tryptamine

0.010

6-Sulfatoxymelatonin

0.020

5-Methoxy-tryptophol

<0.008

5-Methoxy-DL-tryptophan

<0.005

Tryptamine

<0.005

L-Tryptophan

<0.005

N-Acetyl-L-tryptophan

<0.005

5-Hydroxy-tryptamine

<0.005

Melatonin Elisa
Figure 2. Typical melatonin standard curves obtained by RIA and ELISA.

In the ELISA, plasma or serum directly added to the incubation mixture caused a drastic decrease in the optical density measured (data not shown).

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