A number of crop wild relatives can tolerate extreme stress to a degree outside the range observed in their domesticated relatives. However, it is unclear whether or how the molecular mechanisms employed by these species can be translated to domesticated crops. Paspalum (Paspalum vaginatum) is a ...
We previously elucidated principles for designing ideal proteins with completely consistent local and non-local interactions which have enabled the design of a wide range of new αβ-proteins with four or fewer β-strands. The principles relate local bac
Each substrate was tested at various concentrations by incubation with tylosin TE II (1.3 μM) at 30°C in assay buffer (200 mM KPi (pH 7.5), 2.5 mM Tris–HCl (pH 7.5), 50 μM EDTA) in the presence of DTNB (0.2 mM) and 3% (v/v) dimethylsulphoxide to aid substrate solubility....
where they are responsible for the red, purple and blue colours of most flowers, fruits and leaves1,2. Due to their diverse colour expression and many proposed health effects3, their industrial adoption as natural colourants and bioactive compounds in the food, nutraceutical and ...
Cells and tissues (0.3-0.4 g) were ground into fine powder in the presence of liquid nitrogen. Proteins were extracted using 0.7 ml extraction buffer [100 mM Tris, 900 mM sucrose, 10 mM ethylene diamine-tetra-acetic acid (EDTA), 100 mM KCl and 0.4% (v/v) β-mercaptoehtanol, pH ...
The proteasome in eukaryotic cells can degrade a variety of proteins and plays an important role in regulating the cell cycle, cell survival and apoptosis. The proteasome receives much attention as a potential chemotherapeutic target for treatment of a v
Nuclear extracts (5 μg) were preincubated for 15 min at 4°C in 25 μL-binding reaction containing: 10 mM Tris/HCl pH 7.5, 50 mM NaCl, 2.5 mM MgC12, 4% glycerol, 0.5 mM dithiothreitol and 3 μg poly(dI-dC). After adding ~30 pg of labeled DNA, the mixture was incubated for...
(30−50 μg) were separated on 8–12 % SDS-PAGE gels and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, USA). The membranes were then blocked in Tris-buffered saline containing Tween-20 (TBST) with 5% skim milk powder for 1.5 h at room temperature and ...
The pellet was resuspended in the same volume of Protein Loading Buffer (12 mM Tris-HCl pH 6.8 buffer, 5% v/v glycerol, 0.4% SDS, 2.88 mM 2-mercaptoethanol, 0.02% w/v bromophenol blue, 5 mM PMSF), and boiled immediately for 5 minutes. Each lane in the SDS-PAGE gel was...
1c). To exclude false negatives, all investigated GSTs were confirmed to be solubly and functionally produced by SDS–PAGE analysis and an activity test with the generic GST substrate CDNB (5) (Extended Data Fig. 3f,g). Biochemical characterization of PtGSTF8 To characterize the enzymatic ...