The cDNA reactions were diluted to 50 μl in nuclease-free water (Invitrogen Life Technologies), aliquoted and stored at −20 °C till further use. Real time qPCR reactions were carried out in duplicate for each sample using SYBR Premix Ex Taq (Tli RNaseH Plus, DSS Takara ...
A total of 40 microprotein HCPs were identified across the eight antibody products (Fig.4d). Of the 5 sample preparation methods, native digestion protocols resulted in the lowest number of microprotein identifications (Supplementary Fig.8). While we detected two or more peptides from 13 of the...
The nuclease activity of CdtB found in vitro is likely responsible for DNA damage and subsequent G2/M-phase arrest (Lara-Tejero and Galán 2000). However, DNase I function has not yet been fully confirmed, in contrast to the DNA damage that takes place (Méndez-Olvera et al. 2016; Pons...
Flexibility in Mu operator DNA was demonstrated by an unusual supercoil-induced DNA conformation, which was detectable by chemical modification with bromoacetaldehyde or digestion with P1 nuclease. IHF binding adjacent to this site induced dramatic bending of Mu DNA. A topological model we call a ...
Nuclease-free water was used for the blank. The final DNA concentration and purification were determined through fluorometry using a Qubit 2.0 fluorometer (Life Technologies, Grand Island, NY, USA). The total DNA was eluted in 50 μL of elution buffer and stored in a −80 °C freezer ...
0.1% Triton X-100) plus a mammalian cell protease inhibitor mixture (diluted 1:250), 1 mM phenylmethylsulfonyl fluoride, 2.5 mM tosyl phenylalanyl chloromethyl ketone (TPCK), 100 M N-(trans-epoxysuccinyl)-l-leucine 4-guanidinobutylamide (E64), and benzonase nuclease (25 units/ml of ...
nuclease A (SsnA) and the endonuclease A (EndAsuis) led to a reduced bacterial survival after co-cultivation with porcine granulocytes [25,26]. Nevertheless, despite these anti-phagocytic factors,S. suiscannot prevent uptake by neutrophils. Eventually, some bacteria will be phagocytosed and ...
Single-cell suspensions were obtained by flushing the samples through 70 μM strainer, incubated with nuclease (Pierce). Red blood cells were lysed with ACK buffer, and samples stained, according to manufacturer’s instructions. Briefly, cell suspensions were first incubated with 1 μM of 103Rh ...
aureus nuclease and AdsA degrade neutrophil NETs to produce deoxyadenosine and kill macrophages. CAS PubMed PubMed Central Google Scholar Forsgren, A. & Nordström, K. Protein A from Staphylococcus aureus: the biological significance of its interaction with IgG. Ann. NY Acad. Sci. 236, 252...
mRNAs were in vitro transcribed using T7 polymerase purified in-house using linearized plasmid DNA as the template. These mRNAs were capped (Vaccinia Capping System, NEB) and translated using the nuclease-treated rabbit reticulocyte lysate (RRL; Promega). Typical reactions were comprised of 75% v...