Chromatographic separation

A specific and sensitive LC-MS/MS method has been developed for determination of dapaglifozin (DAP) in human plasma. DAP and the internal standard (empaglifozin) were extracted from human plasma by liquid-liquid extraction.

8p vihyuga 04-03-2025 2 1   Download

Recombinant human kynureninase (L-kynurenine hydrolase, EC 3.7.1.3) was purified to homogeneity (60-fold) from Spodoptera frugiperda (Sf9) cells infected with baculovirus containing the kynureninase gene. The purification protocol comprised ammonium sulfate precipitation and several chromatographic steps, including DEAE–Sepharose CL-6B, hydroxyapatite, strong anionic and cationic separations. The purity of the enzyme was determined by SDS/ PAGE, and the molecular mass verified by MALDI-TOF MS. The monomeric molecular mass of 52.4 kDa determined was 99.99% of the predicted molecular mass.

6p research12 01-06-2013 55 5   Download

The disulfide structure of the CRIPTO/FRL-1/CRYPTIC (CFC) domain of human Cripto protein was determined by a combination of enzymatic and chemical fragmentation, followed by chromatographic separation of the fragments, and characterization by mass spectrometry and N-terminal sequencing. These studies showed that Cys115 forms a disulfide bond with Cys133, Cys128 with Cys149, and Cys131 with Cys140.

9p tumor12 20-04-2013 28 3   Download

The rotor subunitc of the A1AOATP synthase of the hyperthermophilic archaeon Pyrococcus furiosuscontains a conserved Na + -binding motif, indicating that Na + is a coupling ion. To experimentally address the nat-ure of the coupling ion, we isolated the enzyme by detergent solubilization from native membranes followed by chromatographic separation tech-niques.

11p galaxyss3 21-03-2013 47 3   Download