Journal: PLoS ONE

Article Title: The CoxD Protein, a Novel AAA+ ATPase Involved in Metal Cluster Assembly: Hydrolysis of Nucleotide-Triphosphates and Oligomerization

doi: 10.1371/journal.pone.0047424

Figure Lengend Snippet: Solubilized CoxD (5.5 mg ml −1 ) was refolded by rapid dilution (50-fold) in ice cold aqueous Tris (20 mM) under magnetic stirring. The solution was immediately adjusted to pH 9.0 (A, dashed line) and subsequently brought to pH 8.0 (A, dotted line) in pH increments of 0.2 per 24 h as specified in . Then the protein was concentrated to 1.04 mg ml −1 by ultra-filtration (A, solid line). The CD spectra of CoxD (0.471 mg ml −1 ; 10 mM KH 2 PO 4 /KOH, pH 8.0) are shown in (B). The CD spectra of CoxD (0.537 mg ml −1 ; 10 mM KH 2 PO 4 /KOH, pH 8.0) in the presence of 0.1 mM MgATP-γ-S are shown in (C). After 2 h of incubation with MgATP-γ-S, excess nucleotides were removed by gel filtration on Sephadex G-25. All data were recorded at 20°C. Raw data are shown in black. The smoothed data used for secondary structure estimation and the back-calculated CD-spectrum based on deconvolution with the CDSSTR algorithm is depicted in red and blue, respectively. For further details refer to .

Article Snippet: Where indicated, sucrose was removed from samples by gel filtration on Sephadex G-25 (PD-10 pre-packed columns, GE Healthcare, Little Chalfont, UK).

Techniques: Filtration, Incubation

Journal: Scientific Reports

Article Title: Anticancer activity of lactoferrin-coated biosynthesized selenium nanoparticles for combating different human cancer cells via mediating apoptotic effects

doi: 10.1038/s41598-023-36492-8

Figure Lengend Snippet: Purification of bovine lactoferrin (bLF). ( A ) Typical elution profile of bLF on CM-Sephadex C50 column. The lactoferrin (●▬●) was eluted at a salt gradient of 0.2–0.6 M NaCl ( ▬ ). All eluted fractions were immunoassayed for bLF confirmation (▲▬▲), and bLF was eluted in the first and second peaks. ( B ) 12% SDS-PAGE for bLF during purification steps. Lane 1 is protein marker, lane 2 is the raw bovine whey, lane 3 is bLF eluted from CM Sephadex C50 column and lane 4 is purified bLF eluted from Sephacryl S200 column.

Article Snippet: At first, bovine whey was applied into the pre-equilibrated CM Sephadex C50 (GE Health care, Sweden) cation exchange column and fractions containing bLF were eluted using 50 mM Tris HCl, pH of 8.0 containing a NaCl gradient of 0.0 to 1.0 M. After that, the contained bLF fractions were applied into a Sephacryl S100 column (5 × 150 mm, GE Healthcare, Sweden), which pre-equilibrated with 50 mM Tris HCl buffer, pH 7.6 and eluted with the same buffer containing 150 mM NaCl.

Techniques: Purification, SDS Page, Marker