Journal: The Journal of biological chemistry

Article Title: Pancreatic stone protein (lithostathine), a physiologically relevant pancreatic calcium carbonate crystal inhibitor?

doi: 10.1074/jbc.272.5.3073

Figure Lengend Snippet: FIG. 2. Analysis of the fractions separated by size exclusion column chromatography. The arrow marks recombinant rat litho- stathine. Lane S represents the selectively concentrated media proteins as loaded onto the column. Proteins were separated by SDS-PAGE (15% acrylamide) and visualized by silver staining. Molecular mass markers are shown at the right (in kDa).

Article Snippet: The fractions were evaluated by SDS-PAGE, and those containing relevant amounts of rat lithostathine were concentrated to a final volume of 9 ml in a Centriprep 10 device and then loaded onto a size exclusion chromatography column (Econo column 25/1200; BioRad) with a P10 matrix (Bio-Rad) and a bed volume of 540 ml.

Techniques: Column Chromatography, Recombinant, SDS Page, Silver Staining

Journal: The Journal of biological chemistry

Article Title: Pancreatic stone protein (lithostathine), a physiologically relevant pancreatic calcium carbonate crystal inhibitor?

doi: 10.1074/jbc.272.5.3073

Figure Lengend Snippet: FIG. 3. Analysis of the fractions eluted from the anion ex- change column. The size exclusion chromatography fractions that were most enriched with lithostathine were pooled and loaded onto a DEAE-Sephacel column (lane S). Proteins were eluted with a gradient of 10–300 mM NaCl. Lane L contains lithostathine purified to apparent homogeneity. Proteins were separated by SDS-PAGE (15% acrylamide) and visualized by silver staining. Molecular mass markers are shown at the right (kDa).

Article Snippet: The fractions were evaluated by SDS-PAGE, and those containing relevant amounts of rat lithostathine were concentrated to a final volume of 9 ml in a Centriprep 10 device and then loaded onto a size exclusion chromatography column (Econo column 25/1200; BioRad) with a P10 matrix (Bio-Rad) and a bed volume of 540 ml.

Techniques: Size-exclusion Chromatography, Purification, SDS Page, Silver Staining

Journal: The Journal of biological chemistry

Article Title: Pancreatic stone protein (lithostathine), a physiologically relevant pancreatic calcium carbonate crystal inhibitor?

doi: 10.1074/jbc.272.5.3073

Figure Lengend Snippet: FIG. 4. Immunodetection of rat lithostathine. Proteins contained in the fractions of the anion exchange chromatography were separated by SDS-PAGE (see Fig. 3) and transblotted onto a polyvinylidene diflu- oride membrane. Baculovirus expressed rat lithostathine was detected using anti-rat lithostathine antibodies. L and S as in Fig. 3. Molecular mass markers are shown at the right (in kDa).

Article Snippet: The fractions were evaluated by SDS-PAGE, and those containing relevant amounts of rat lithostathine were concentrated to a final volume of 9 ml in a Centriprep 10 device and then loaded onto a size exclusion chromatography column (Econo column 25/1200; BioRad) with a P10 matrix (Bio-Rad) and a bed volume of 540 ml.

Techniques: Immunodetection, Chromatography, SDS Page, Membrane

Journal: Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry

Article Title: Dual Effect of Heparin on Fe 2+ Induced Cardiolipin Peroxidation. Implications to Peroxidation of Cytochrome c Oxidase Bound Cardiolipin ‡

doi: 10.1007/s00775-013-1019-z

Figure Lengend Snippet: Upper panel – silicic acid HPLC chromatogram. The numbers (I, II and III) designate the randomly chosen areas where spectra were analyzed. Lower panels ( b, c and d ) represent spectra extracted from the randomly chosen areas (I, II and III) of silicic acid HPLC. Solid line is CL extracted from 5 μM CcO; dotted line is CL extracted from 5 μM CcO incubated with 5 mM tert-BOOH and 50 μM Fe2+; dashed line is CL extracted from 5 μM CcO incubated with 5 mM tert-BOOH, 50 μM Fe2+ and 100 μM heparin.

Article Snippet: The silicic acid HPLC column (5μm Radial Pac Resolve Silica cartridge, 0.8 cm × 10 cm) was purchased from Waters Corporation, Inc. All other chemicals were reagent grade.

Techniques: Incubation

Journal: Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry

Article Title: Dual Effect of Heparin on Fe 2+ Induced Cardiolipin Peroxidation. Implications to Peroxidation of Cytochrome c Oxidase Bound Cardiolipin ‡

doi: 10.1007/s00775-013-1019-z

Figure Lengend Snippet: CcO (5μM) solubilized in 50 mM Tris-SO4 buffer, pH 7.4, containing 2 mM dodecyl maltoside and 2 mM sodium cholate was incubated for 60 min at room temperature with: (1) no additions; (2) 5 mM tert-BOOH ± 100 μM heparin ; (3) 5 mM tert-BOOH + 50 μM Fe2+; (4) 5 mM tert-BOOH + 50 μM Fe2+ + 100 μM heparin; (5) 50 μM Fe2+ ; (6) CcO + 50 μM Fe2+ + 100 μM heparin. The reaction was stopped by dilution and the activity measured spectrophotometrically with reduced cytochrome c as a substrate. These results are representative of three independent experiments. The error in determination of molecular activity was ±2.6%.

Article Snippet: The silicic acid HPLC column (5μm Radial Pac Resolve Silica cartridge, 0.8 cm × 10 cm) was purchased from Waters Corporation, Inc. All other chemicals were reagent grade.

Techniques: Incubation, Activity Assay