SF samples were diluted 4-fold in calibrator diluent or assay buffer and were measured in duplicate.
The interactions were visualized with 0.05% 2,2 azino di-3-etilbenziatioazolina sulphonic acid (ABTS) (Sigma) in 100 mM citrate buffer, and were measured (optical density at 405 nm) in a spectrophotometer (Multiskan Bichromatic, Labsystems, Finland).
The free iron oxide (Fed) was extracted by a Dithionite citrate system buffered with sodium bicarbonate (DCB) and was measured with atomic absorption spectrometer (AAS).
For analysis, cells were resuspended in 500 ml of buffer and fluorescence was measured using the FACS Calibur BD Biosciencee) and analysed using WinMIDI 2.8 software.
Kidney slides were mechanically lysed in RIPA buffer and fluorescence was measured in a fluorometer (Turner) with excitation of 450 nm and emission of 530 nm.
The protein was extracted with lysis buffer and concentration was measured using a microplate reader.
Pellets were resuspended in 100 μL of 1× PBS– buffer, and fluorescence was measured at 665 nm.
Cells were then washed two more times with PBS buffer and fluorescence was measured by FACSCalibur using CellQuest software (BD Biosciences).
Hundred microlitres of each serum was mixed with assay buffer and absorbance was measured at 420 nm using a Synergy 2 ELISA plate reader (Bio-Tek).
After three washes, phosphatase substrate (Roche) was added at 1 mg/ml in substrate buffer and absorbance was measured at 415 nm.
