Search results for "acrylamide"
showing 10 items of 485 documents
Construction of an agglutination tool: recombinant Fab fragments biotinylated in vitro.
2009
The pComb3H vector system is used for constructing and panning recombinant antibody libraries. It allows for expression of monovalent Fab fragments, either on the surface of M13 phage, or in the form of soluble proteins secreted into the periplasmic space of bacteria. We constructed a modified pComb3H vector containing cDNA encoding for a 23-amino acid fragment of the Escherichia coli biotin carboxy carrier protein (BCCP), which is an acceptor sequence for biotinylation. The vector was used to express the Fab fragment recognizing human glycophorin A. The purified Fab fragment containing this biotin acceptor sequence was effectively biotinylated in vitro using biotin ligase (BirA). The speci…
Mobility of Acetylated Histones in Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis
1999
Abstract We describe an altered mobility for acetylated histone isoforms in sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Isoforms of histones H3 and H4 with a higher acetylation degree have a slightly faster electrophoretic mobility. Since acetylation neutralizes the positive charge of the e-amino group of lysine, without significantly changing the molecular mass of the protein, the acetylation-dependent mobility shift could be explained by the increase of the net negative charge of the SDS–histone complexes. A possible consequence of this differential mobility for the acetylation site determination by protein microsequencing from SDS gels is discussed.
Exposure to malondialdehyde induces an early redox unbalance preceding membrane toxicity in human erythrocytes.
2002
This work investigated the oxidative injury to human red blood cells (RBCs) by the exposure to exogenous malondialdehyde (MDA), in a physiological environment. When a 10% RBC suspension was incubated in autologous plasma, in the presence of 50 microM MDA, 30% of MDA entered into the cells. A time-course study showed that MDA caused early (30-120 min) and delayed (3-18 h) effects. MDA caused a fast depletion of reduced glutathione, and loss of the glucose-6-phosphate dehydrogenase activity, followed by a decrease of HbO2. Accumulation of methemoglobin, and formation of small amounts of hemichrome were later evident. Also, an HbO2-derived fluorescent product was measured in the membrane. The …
Electrophoretic separation of a class of nucleosomes enriched in HMG 14 and 17 and actively transcribed globin genes.
1980
Monomer nucleosomes from chick erythrocytes can be fractionated according to their electrophoretic mobility in (comparatively) high salt acrylamide gels. We show that the fractionation is based predominantly on differences in charge. The monomer heterogeneity persists even when the nucleosomes are trimmed down to 145 bp with Exo III or when H1 and H5 are removed. The slowest migrating monomers are associated with HMG 14 and 17; however, we do not believe that these proteins are entirely responsible for the altered mobility since the nucleosome heterogeneity persists even after removal of HMG 14 and 17. The DNA associated with the HMG 14 and 17 containing nucleosomes is shown to be enriched …
Neutral deoxyribonucleases of HeLa S3 cells: electrophoretic separation, characterization, substrate specificity and mode of action.
1980
Extracts of HeLa S3 cells were electrophoresed on polyacrylamide gels; gel slices were eluted and the eluates were assayed for DNase activities against native and denatured DNA substrates in the presence of MgCl2 or Na2EDTA. Aliquots of each eluate were also assayed for their ability to nick the circular supercoiled PM2 phage DNA to distinguish endonucleases from exonucleases. Peaks of endonuclease activities were characterized as forming 3′-phospho-oligonucleotides or 5′-phospho-oligonucleotides by the use of oligonucleotides produced by these enzymes as substrates for the 5′-phosphate-specific snake venom exonuclease. The total activity of DNases in gel eluates was much higher than that i…
Development of Antioxidant Protein Extracts from Gilthead Sea Bream (Sparus aurata) Side Streams Assisted by Pressurized Liquid Extraction (PLE)
2021
The pressurized liquid extraction (PLE) technique was used, for the first time, to obtain protein extracts with antioxidant activity from side streams (muscle, heads, viscera, skin, and tailfins) of gilthead sea bream (Sparus aurata) in order to give added value to these underutilized matrices. Extraction conditions previously optimized for sea bass (Dicentrarchus labrax) side streams were applied. Protein recovery percentages were 22% (muscle), 33% (heads), 78% (viscera), 24% (skin), and 26% (tailfins), which represented an increase of 1.2–4.5-fold compared to control samples (extraction by stirring). The SDS-PAGE profiles revealed that PLE-assisted extraction influenced protein molecular …
Effect of lactic acid fermentation of lupine wholemeal on acrylamide content and quality characteristics of wheat-lupine bread
2013
The effect of supplementing wheat flour at a level of 15% with lupine (Lupinus angustifolius L.) wholemeal fermented by different lactic acid bacteria on acrylamide content in bread crumb as well as on bread texture and sensory characteristics was analysed. The use of fermented lupine resulted in a lower specific volume and crumb porosity of bread on an average by 14.1% and 10.5%, respectively, while untreated lupine lowered the latter parameters at a higher level (30.8% and 20.7%, respectively). The addition of lupine resulted in a higher by 43.3% acrylamide content compared to wheat bread (19.4 µg/kg dry weight (d.w.)). Results showed that acrylamide was significantly reduced using proteo…
Microbial inactivation and evaluation of furan formation in high hydrostatic pressure (HHP) treated vegetable-based infant food.
2017
Abstract The inactivation of pathogenic and spoilage bacteria as well as the formation of food processing contaminants (e.g. acrylamide, furan, etc.) in infant foods is of utmost importance for industry, consumers as well as regulatory bodies. In this study, the potential of high hydrostatic pressure (HHP) for microorganism inactivation including total mesophilic aerobic bacteria (TMA) and total yeasts and molds (TYM) at equivalent processing conditions, as well as its effects on furan formation in vegetable-based infant food was evaluated. The process parameters evaluated were combinations of pressures (200, 300, and 400 MPa), temperatures (25, 35, and 45 °C), and treatment times (5, 10, a…
TTAS a New Stilbene Derivative that Induces Apoptosis in Leishmania Infantum
2012
Leishmania parasites are able to undergo apoptosis (programmed cell death), similarly to mammalian cells. Recently it was demonstrated in vitro the anti-leishmanial effect of some natural and synthetic stilbenoids including resveratrol and piceatannol. In this study we evaluated the Leishmanicidal activity of a pool of stilbene derivatives which had previously shown high apoptotic efficacy against neoplastic cells. All the compounds tested were capable to decrease the parasite viability in a dose-dependent manner. Trans-stilbenes proved to be markedly more effective than cis-isomers. This was different from that observed in tumor cells in which cis-stilbenes were more potent cytotoxic agent…
Modified apparatus for voltage gradient gel electrophoresis.
1999
We built a modified version of voltage gradient gel electrophoresis system to correct distortions in nucleic acids electrophoretic migration patterns occurring at the edges of the gel when the original voltage gradient apparatus is used. The new device allows correct fractionation of nucleic acids also when electrophoresis is performed at high voltages.