aRNA was fragmented using 5x fragmentation buffer and the total amount of fragmented biotinylated aRNA used per chip was 6.5 μg.
After 48 h, samples were lysed using 1x Passive lysis buffer, and the amount of firefly luciferase and Renilla luciferase was quantified using the Dual-Luciferase Reporter Assay System Promega Corporationn, Masison, WI), as we described recently [15].
Colospheres, spheroids and xenograft tissue were lysed with zymogram sample buffer and the amount of protein was estimated using the Bradford method (Biorad Dc Protein Assay; Bio-Rad Laboratories).
Protein bound to the beads was washed three times in lysis buffer and the amount of bound fusion protein was estimated using Coomassie-stained SDS gels.
After immunoprecipitation, beads were washed three times with lysis buffer and the amount of HCF101 Myc-associated radioactivity was measured by scintillation counting.
Tissue samples or myofibroblasts were homogenized in ice-cold lysis buffer, and the amount of protein was determined by a protein assay (Bio-Rad Laboratories).
The ratio of oral secretion to Mes-NaOH buffer and the amount of OS solution added to mechanical injured (MI) tomato plants were as reported in previous studies [ 3, 6, 10, 11, 32].
At the end of incubation, liposomes were washed 5 times with the same buffer by centrifugation, and the amounts of bound tTG were quantified in a gamma counter.
Finally, Au-citr CTAB -TbADH bioconjugAu-citr CTAB -TbADHAu-citr CTAB -TbADHhosphate bioconjugate.0, and the amount of immobilized enzyme was calculated from the difference in the protein amount before and after immobilization process.
For Western blotting, cells were prepared by lysing in Laemmli sample buffer and equal amounts of protein, according to the cell count, were resolved by 12% SDS PAGE.
