Immunoenzymatic method (ELISA) for quantifying the iron deficiency in phytoplankton using photosynthetic proteins.

摘要

Immunoenzymatic method (ELISA) for quantifying the iron deficiency in phytoplankton using photosynthetic proteins - Quantitative immunoenzymatic method (ELISA) for detecting and quantifying photosynthetic redox proteins in green algae that are affected by the availability of iron in the environment. This method consists of several steps: a) depositing a phytoplankton anti-protein specific antibody on the craters of a plate made of polyethylene or other material that can fix proteins: b) washing to eliminate the antibodies that have not been fixed to the plate: c) incubating with a solution of a protein foreign to the test and then washing: d) applying, to each crater of the plate, the phytoplankton extract from the water to be analysed, or other known concentration patterns: e) washing to eliminate the proteins that have not reacted to the antibodies: f) incubating each crater of the plate with phytoplankton anti-protein antibodies (directed against the same protein than the one used in step "a"), conjugated with peroxidase or alkaline phosphatase and diluted in a buffer at room temperature: g) washing to eliminate the antibodies that have not reacted: g) incubating each crater with a substrate appropriate for the enzyme that was used for its development: i) after some time has passed, measuring the absorbance of each crater. The proteins against which the antibodies are induced are the flavo-oxidase and ferredoxin of the phytoplankton organism itself that perform the same function, depending on the bioavailability of iron in the culture medium.