To gain insights on the interaction of collagen and minerals at the nanoscale, bone samples were deproteinized or demineralized.
After reversing crosslinks, the samples were deproteinized and phenol-chloroform extracted, then DNA was ethanol precipitated using yeast tRNA as a carrier.
In particular, urine samples were deproteinized by 1∶5 dilution in 50% acetonitrile with 2 mM debrisoquine sulfate and 30 mM 4-nitrobenzoic acid as internal controls.
Plasma samples were deproteinized by 1∶40 dilution in 66% acetonitrile containing 2 mM debrisoquine sulfate and 30 mM 4-nitrobenzoic acid as internal controls.
Samples were deproteinized with an equal volume of 10% metaphosphoric acid, centrifuged, and the supernatant was analyzed for GSH and GSSG concentrations using a spectrophotometer at 412 nm over 3 min. Concentrations were calculated by comparison to GSSG standards and normalized to protein content.
The blood samples were deproteinized with ZnSO4 and Ba(OH 2, dried to remove 3H2O, re-suspended in water, and counted in scintillation fluid (BioSafe II, Research Product International Corp., Mount Prospect, IL) on dual channels to provide separation of 3H and 14C counts for the determination of plasma [2 3H]glycerol and [1 14C]glucose, respectively.
Samples were deproteinized using acetonitrile.
Plasma samples were deproteinized with trichloroacetic acid.
The samples were deproteinized as described [ 56, 57], and fractionated through 0.8% AGE with Ethidium Bromide.
The plasma samples were deproteinized using acetonitrile at a final concentration of 80% before measurement.
