The LSPR-based sensors were incubated overnight and then rinsed with PBS buffer, and extinction spectra were collected in PBS buffer to quantify the miR-10b levels.
Cells were washed in phosphate buffered saline (PBS) and whole cell lysates were collected in ELB buffer (50 mM HEPES (pH 7), 250 mM NaCl, 0.1% Nonidet P-40, 5 mM EDTA, 10 mM NaF, 0.1 mM Na3VO4, 50 µM ZnCl2, supplemented with 0.1 mM PMSF, 1 mM DTT, and a mixture of protease inhibitors, phosphatase inhibitors were not added).
In brief, animals were collected in M9 buffer, frozen in liquid nitrogen and boiled in SDS sample buffer für 10 minutes.
7 Buccal or fibroblast cells were collected in extraction buffer (Abcam, Cambridge, MA) and mixed with sodium dodecyl sulfate (SDS) sample buffer.
Two-milliliter samples of whole blood were collected in buffered sodium citrate, and high molecular weight genomic DNA was obtained from the peripheral blood leukocyte fraction (QIAamp Blood Tissue Kit, Qiagen, Valencia CA).
Animals were collected in M9 buffer and frozen in liquid nitrogen.
HEK293 cells were collected in lysis buffer, sonificated, and insoluble material was removed by centrifugation.
Proteins were desalted by passing through PD-10 gel filtration columns and were collected in appropriate buffer before each experiment and assays.
CD3+ T cell lysates were collected in lysis buffer (10 mM Tris,pH 7.4, 1 mM sodium orthovanadate, 1% SDS), sonicated and frozen at −80°C.
The resulted lysates were collected in Laemmli buffer and subjected to 14% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the separated proteins were transferred to polyvinylidene difluoride membrane (Bio-Rad).
