To minimize artificial oxidation, 2 µg of butylated hydroxytoluene (BHT) was immediately added to the diluted plasma, which was then incubated at 37 °C for 10 min with 10 µl of proteinase K to eliminate non-exosomal plasma proteins. At the end of the proteinase K treatment,...
most of these affecting the central and/or peripheral nervous systems. Patients typically show developmental delay or intellectual disability, seizures, neuropathy, and metabolic abnormalities in multiple organ systems3. Adding the correct sugar chains (glycans) to proteins and lipids significantly impacts ...
Innovations in high-throughout sequencing approaches are being marshaled to both reveal the composition of the abundant and heterogeneous noncoding RNAs th
Vertebrate early embryogenesis is initially directed by a set of maternal RNAs and proteins, yet the mechanisms controlling this program remain largely unknown. Recent transcriptome-wide studies on RNA structure have revealed its pervasive and crucial roles in RNA processing and functions, but whether ...
DNA from mice tail snips was extracted at weaning (21 days) using Genotyping buffer containing 0.1M Tris PH7.5, 0.05M EDTA, 0.2% (v/v) SDS, 0.2M NaCl and proteinase K solution incubated overnight at 55 °C and DNA precipitated with isopropanol. Dried DNA pellet was then resuspended in ...
Part of the CFS was buffered with sterile 0.5 M NaOH solution in order to restore the initial pH of the medium (pH 7.80 ± 0.20) and obtain a neutralized cell-free supernatant (CFSn). Subsequently. a fraction of CFSn was treated with proteinase K (Sigma-Aldrich) as previously...
Two different DNA fragments, deriving from the P22-related bacteriophage LP7 and from the right IS element of transposon Tn10, have been identified which were recognized by the P22-packaging apparatus as pac-like signals. Their nucleotide sequences were compared with the known sequence of P22 gen...
A total of 40 μL of the remaining diluted plasma was mixed with 160 μL PBS to obtain a final volume of 200 μL and was used directly for DNA isolation without filtering. For DNA isolation, 20 µL Proteinase K and 200 µL AL buffer (from DNA isolation kit) were added to the...
Cell fractionation experiments indicated that the bulk of the PDIP38 in cells was in a mitochondrial pellet, and resistant to proteinase K digestion until the membranes were solubilized with Triton X-100; a smaller amount was present in the nuclear fraction. Immunofluorescence studies of endogenous ...
Disruption of β-amyloid (Aβ) homeostasis is the initial culprit in Alzheimer’s disease (AD) pathogenesis. Astrocytes respond to emerging Aβ plaques by altering their phenotype and function, yet molecular mechanisms governing astrocytic response and t