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Ferrozine‐Based Assay for Determining Iron Content in Insect Cells
Author(s) -
Castaneda Mirella,
ALOgaidi Hussein,
Gorman Maureen J.,
Ragan Emily J.
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.636.2
Subject(s) - ferrous , chemistry , detection limit , ferric , hemolymph , colorimetric analysis , ferric iron , microbiology and biotechnology , biochemistry , chromatography , biology , inorganic chemistry , organic chemistry
We are optimizing a ferrozine‐based assay to analyze the iron content in Drosophila S2 cells. We hypothesize that putative ferric reductases CG1275 or CG8399 may be involved in reducing iron in the hemolymph and allowing uptake into cells. We have modified a ferrozine‐based assay, in which ferrozine complexes with ferrous ions in solution and forms a stable magenta complex species that can be detected at 562 nm. Using this method with an Ultraspec II spectrophotometer we obtained a limit of detection of 0.30 μg/mL. This method provides a reliable, inexpensive and simple way of analyzing and quantifying the iron content within the S2 cells, allowing us to analyze iron content after RNA interference experiments to knock down CG1275 and CG8399. Support or Funding Information NSF IOS‐1656407 This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .