Gold Nanomaterials at Work in Biomedicine *
Valerio Voliani in Nanomaterials and Neoplasms, 2021
As one of the most abundant plasma proteins, bovine serum albumin (BSA) is widely used in an array of applications such as sensing, self-assembly, and imaging [81]. Ying and coworkers demonstrated the use of BSA as a template for the synthesis of Au clusters [82], similar to what was reported for the Ag clusters [78]. As shown in Fig. 5.7B, upon the addition of Au(III) ions into an aqueous BSA solution, the protein molecules sequestered Au(III) ions and entrapped them. When the pH value was increased to 12, the reduction power of BSA was enhanced and the entrapped ions would go through progressive reduction to generate Au clusters. Based on the photoluminescence property and the spherical jellium model, the BSA-stabilized Au clusters were proposed to contain a Au25 core, which was consistent with the MALDI-MS data. Due to the excellent biocompatibility and abundant functional groups of BSA, this class of Au clusters remains highly promising for various applications in biomedicine.
The Production of Superoxide by Cultured Macrophages
Robert A. Greenwald in CRC Handbook of Methods for Oxygen Radical Research, 2018
The cells remaining in the dish are washed gently three times with buffer. Copper tartrate reagent8 is added directly to the dish, and the protein content of the dish is determined by the method of Lowry et al.8 using bovine serum albumin (BSA) as standard. The dishes used as blanks give a significant reaction, and this value must be subtracted from that of the dishes containing cells. The extent of O2− release can be corrected for the protein content of each individual dish if one is certain that cells are not dislodged during the incubation or subsequent washes. Alternatively (and preferably), if macrophage adherence is uniform from dish to dish, the mean of the protein content of 3 to 5 dishes washed free of nonadherent cells but not carried through the assay can be used for expression of the extent of O2− release in individual dishes. Release of O2− can also be expressed on the basis of cell number, using a conversion factor that equates cell protein and number4 or, preferably, an actual determination of cell number by counting nuclei of disrupted cells.9
Selection of Material for Dialysis Membrane
Sirshendu De, Anirban Roy in Hemodialysis Membranes, 2017
The bicinchoninic acid (BCA) protein assay11 was used to estimate the total protein content in this method. NIH3T3 cells were seeded on samples on days 3 and 5. PBS-rinsed cell-seeded samples were incubated with BCA working solution (50 parts of BCA reagent with 1 part of 4% copper sulfate pentahydrate, green-colored solution) at 37°C for 30 minutes. The mechanism involved in this is the reduction of free amino acids, which form a crimson-colored complex with BCA. The concentration of this was measured by measuring absorbance at 562 nm on a microplate reader (Recorders and Medicare Systems, India). The standard protein concentration curve was plotted with a known concentration of bovine serum albumin (BSA).
Synthesis, cytotoxicities, and carbonic anhydrase inhibition potential of 6-(3-aryl-2-propenoyl)-2(3H)-benzoxazolones
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2019
Sinan Bilginer, Halise Inci Gul, Feyza Sena Erdal, Hiroshi Sakagami, Serkan Levent, Ilhami Gulcin, Claudiu T. Supuran
The purification of cytosolic CA isoenzymes (CA I and CA II) was previously described with a simple one-step method by a Sepharose-4B-L tyrosine-sulphanilamide affinity chromatography42,53,55–58. The protein quantity in the column effluents was determined spectrophotometrically at 280 nm. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was applied with a Bio-Rad Mini Gel system Mini-PROTEIN system (Bio-Rad Laboratories, Inc., Shanghai, China) after purification of both CA isoenzymes. Briefly, it was performed in acrylamide for the running (10%) and the stacking gel (3%) contained SDS (0.1%), respectively. The increase in absorbance of the reaction medium was spectrophotometrically recorded at 348 nm. Also, the quantity of protein was determined at 595 nm according to the method as described previously55–60. Bovine serum albumin was used as a standard protein. The IC50 values were obtained from activity (%) versus compounds plots. For the calculation of Ki values, three different concentrations were used. The Lineweaver–Burk curves were drawn and calculations were realised as before55–60.
Exploring new structural features of the 4-[(3-methyl-4-aryl-2,3-dihydro-1,3-thiazol-2-ylidene)amino]benzenesulphonamide scaffold for the inhibition of human carbonic anhydrases
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2019
Simona Distinto, Rita Meleddu, Francesco Ortuso, Filippo Cottiglia, Serenella Deplano, Lisa Sequeira, Claudia Melis, Benedetta Fois, Andrea Angeli, Clemente Capasso, Rossella Angius, Stefano Alcaro, Claudiu T. Supuran, Elias Maccioni
The protein quantity in the column effluents was determined spectrophotometrically at 280 nm. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was applied with a Bio-Rad Mini Gel system Mini-PROTEINVR system (Hercules, CA), Bio-Rad Laboratories, Inc., China after purification of both CA isoenzymes. Briefly, it was performed in acrylamide for the running (10%) and the stacking gel (3%) contained SDS (0.1%), respectively. Activities of CA isoenzymes were determined according to a method by Verporte et al.57. The increase in absorbance of the reaction medium was spectrophotometrically recorded at 348 nm. Also, the quantity of protein was determined at 595 nm according to the Bradford method58. Bovine serum albumin was used as standard protein. The IC50 values were obtained from activity (%) versus compounds plots59. For calculation of KI values, three different concentrations were used. The Lineweaver–Burk curves were drawn and calculations were realised59. The biological data are reported in Table 3.
PU.1 interaction with p50 promotes microglial-mediated inflammation in secondary spinal cord injury in SCI rats
Published in International Journal of Neuroscience, 2023
Mingchen Yu, Yiqing Ou, Hongmei Wang, Weidong Gu
The sections to be studied (sham operation, 3 days post-SCI) were heated in a 37 °C oven for 2 h, followed by two 5-min washes with 0.01 M PBS. Next, the sections were blocked with PBS containing 5% bovine serum albumin for 2 h and then permeated with 0.1% Triton X-100/PBS for 20 min. The sections were then incubated with the primary monoclonal antibody PU.1 (rabbit, 1:100; Abcam) or antibodies of different cellular markers, such as NeuN (neuron marker, mouse, 1:500; Chemicon, Temecula, CA, USA), GFAP (astrocyte marker, mouse, 1:100; Sigma), and Iba-1 (cytoplasmic marker of microglia, mouse, 1:100; Serotec, Oxford, UK), for 12 h. The antibodies were prepared in 1% bovine serum albumin. After incubation, the sections were washed 3 times with PBS, 10 min per wash, and then incubated with the secondary antibody (CY3-Donkey or FITC-Donkey, 1:1000; Jackson Laboratory, Bar Harbor, ME, USA) in the dark at RT for 2 h. The stained sections were observed with a Leica fluorescence microscope or Leica confocal microscope.
Related Knowledge Centers
- Isoelectric Point
- Ph
- Signal Peptide
- Blood Plasma
- Serum Albumin
- Cohn Process
- Blood Protein
- Salt
- N-Terminus
- Molar Absorption Coefficient