Protoporphyrin 9 pp

HA with a mean molecular weight of 30,000 (polydispersity index of 1.212) was purchased from MRC polysaccharide Corp., Ltd (Toyama, Japan). RPMI 1640 medium, 4-dimethylaminopyridine (DMAP) isoflurane, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Tween 20, sodium dodecyl sulfate (SDS) and zinc acetate were purchased from Wako pure chemical (Osaka, Japan). Protoporphyrin IX (PP) and cetyltrimethylammonium bromide (CTA) were from Sigma-Aldrich Chemical (St. Louis, MO, USA). Water-soluble carbodiimide (WSC) was purchased from Dojindo Laboratory, Kumamoto, Japan. Additionally, 2,2,6,6-Tetramethyl-4-piperidone (4-oxo-TEMP) was purchased from Tokyo Chemical Industry (Tokyo, Japan). Esterase from porcine liver was purchased from Roche Diagnostics GmbH (Mannheim, Germany). The other reagents and solvents of a reagent grade were from commercial sources and were used without further purification.

Swine hematite (SH) from Becton & Dickinson was used as positive control and protoporphyrin IX (PP) from Sigma-Aldrich as negative control. These molecules were characterized using the same analytical techniques and were selected to differentiate the presence and absence of iron, in the SH and PP analyses, respectively. That compared to the use of control molecules of Hz and β-Hz (17 , 26 ) it is only possible to analyze the difference due to the formation of the dimer. All the described reagents were analytical grade and the solutions were prepared using deionized water with 18 M MΩ.