H2O2 release into the incubation buffer was quantified by the horseradish peroxidase-catalyzed oxidation of 10-acetyl-3,7-dihydroxy-phenoxazine 10-acetyl-3,7-dihydroxy-phenoxazine 10-acetyl-3,7-dihydroxy-phenoxazine 10-acetyl-3,7-dihydroxy-phenoxazine 10-acetyl-3,7-dihydroxy-phenoxazine
Cells were harvest in caspase assay buffer, and proteins were quantified by Bradford assay.
Cells were lysed in RIPA buffer, and protein concentrations were quantified by using Bradford reagent.
Cell lysates were prepared with RIPA buffer and the total proteins were quantified by BCA assay as mentioned above.
BMMSCs were harvested in RIPA lysis buffer (Beyotime Institute of Biotechnology) and whole-cell protein extracts were quantified by a BCA assay, separated on SDS-PAGE 8%-12%, and then transferred to PVDF membranes (Millipore, Billerica, MA, USA).
After five washes in lysis buffer, GST fusion proteins bound to the beads were quantified by performing a Bradford assay (Bradford Reagent, SIGMA), aliquoted and kept at − 80 °C in lysis buffer containing 10% glycerol (binding buffer).
Cells were harvested in RIPA lysis buffer (Beyotime Co .. Whole-cell protein extracts were quantified by the BCA assay, separated on 12% SDS-PAGE and then transferred to PVDF membranes (Millipore, Billerica, MA, USA) and blocked with 5% nonfat milk powder in PBST for 3 h.
After Triton-X100 exchange, incubation overnight in developing buffer and staining with Coomassie Blue, bands of lysis were quantified by densitometry and expressed as arbitrary optical units.
The adherent viable cells were quantified by MTS after rinsing in excess buffer solution.
RhoA levels were quantified by running bead/protein complexes in loading buffer containing 0.1 M DTT.
