The reaction mixtures contained, in a final volume of 1.2 ml, the following reagents at the final concentrations stated: deoxyribose (2.8 mM), ferric chloride Fe[Cl.sub.3] (25 [micro]M), nitrilotriacetic acid NTA (100 [micro]M) (NTA and [Fe.sup.3+] ions are pre-mixed at the ratio given prior to the addition of deoxyribose), phosphate buffer ([K.sub.2]HP[O.sub.4]-K[H.sub.2]P[O.sub.4]) pH 7.4 (10 mM), different concentrations of CE, EAF and BF (0.1-100 [micro]g [ml.sup.-1]) and hydrogen peroxide [H.sub.2][O.sub.2] (2.8 mM).
Dimethyl sulfoxide (DMSO), thiobarbituric acid (TBA), trichloroacetic acid (TCA), mannitol, butylated hydroxytoluene (BHT), t-butyl-peroxide, deoxyribose, ferric chloride, nitrilotriacetic acid (NTA), hydrogen peroxide, superoxide dismutase, phenazine methosulphate, nicotinamide nucleotide diphosphate and nitro blue tetrazolium (NBT) were purchased from Sigma.
The complexes of citrate and
nitrilotriacetic acid. J Biol Chem 1967;242: 2810-5.
Dextran sulfate sodium salt and CNBr-activated Sepharose 6B were obtained from Amersham Biosciences;
nitrilotriacetic acid trisodium salt monohydrate (NTA) was from Fluka; bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane (Bis-Tris), calcium chloride dehydrate (Ca[Cl.sub.2] 2 [H.sub.2]O), and iron(III) chloride hexahydrate (Fe[Cl.sub.3] 6 [H.sub.2]O) were from Sigma-Aldrich (Stockholm, Sweden); bilirubin was from VWR International; and rabbit anti-transferrin antibodies (Q327) were from Dakopatts A/S.
The reaction mixture contained the following reagents (final concentrations in brackets) in a final volume of 1.2 ml: deoxyribose (2.8 mM), ferric chloride [FeCl.sub.3] (25 [micro]M),
nitrilotriacetic acid (NTA; 100 [micro]M) (NTA and [Fe.sup.3+] ions are premixed at the ratio given, prior to the addition of deoxyribose), phosphate buffer ([K.sub.2][HPO.sub.4]-[KH.sub.2][PO.sub.4]) pH 7.4 (10 mM), different concentrations of CE, fractions, sub-fractions and isolated compounds (0.1 to 100 [micro]g*[ml.sup.-1]) and hydrogen peroxide ([H.sub.2][O.sub.2] 2.8 mM).
[4] Nonstandard abbreviations: NTBI, non-transferrin-bound iron; NTA,
nitrilotriacetic acid; AAS, atomic absorption spectrophotometry; FeNTA, ferric
nitrilotriacetic acid; PBS, phosphate-buffered saline; and PAGE, polyacrylamide gel electrophoresis.