A protocol for high-throughput affinity determinations using a protein microarray and fluorescently labeled synthetic peptides was published by Kaushansky et al. [ 46].
The protein concentration was measured using a Cayman kit (Protein Determination Reagent item no. 704004 and Protein Determination BSA standard item no. 704 003), which is based on the Bradford method.
Protein quantification was performed using Bio-Rad protein determination reagent.
Supernatants were subjected to protein determinations using a DC protein assay kit (Bio-RAD Laboratories, HerCAles, Caccordinging to the manufacturer's instructions.
Protein concentrations were determined using the DC protein determination kit (BioRAD) according to manufacturer's protocol and samples volumes were adjusted for loading.
After centrifuge, the supernatant was collected and used for protein determination with a BCA Protein Assay Kit (Pierce).
The supernatant of the homogenate was used for protein determination with a BCA Protein Assay Kit (Pierce, IL, USA) and electrophoresis.
After centrifuge, the supernatant of the homogenate was collected and used for protein determination with a BCA Protein Assay Kit (Pierce, Rockford, IL, USA).
The pellets of the acidic extract were dissolved in 50 μl of NaOH (0.1M) and used for protein determination at a dilution of 1 10 (BCA Assay; Pierce).
After protein content determination using a DC Protein Assay kit (Bio-Rad Laboratories, HerCAles, CA, USA) and subsequently incubation with dilute solution (1 : 1000) of primary antibodies including anti-Bax, anti-Bcl-2, anti-caspase3, and anti-actin (Santa Cruz Biotechnology Inc, Santa Cruz, CA).
