Adipokine proteome profiler kit

The effect of coconut water vinegar on the secreted cytokines from the adipose tissue was analysed using an adipokine proteome profiler kit (R&D Systems, Minneapolis, USA). In brief, the membranes were blocked using a blocking buffer at room temperature for 1 h. Concurrently, the protein samples were incubated with the detection antibody at room temperature for 1 h. Next, the protein samples coupled with the detection antibody were added to the membrane and incubated at 4°C overnight. The next day, the membranes were washed three times using a washing buffer and incubated with streptavidin–horseradish peroxidase for 30 min. Finally, the membranes were washed a further three times before the substrate was added to develop a chemiluminescence condition. The membrane was then viewed using a ChemiDoc XRS (Bio-Rad, USA).

The effect of pineapple vinegar on secreted cytokine from adipose tissue was analyzed using an adipokine proteome profiler kit (R&D System, USA). In brief, the membranes were blocked using blocking buffer at room temperature for 1 hour. Concurrently, the protein samples were incubated with the detection antibody at room temperature for 1 hour. Next, the protein samples, coupled with the detection antibody, were added to the membrane and incubated at 4°C overnight. The next day, the membranes were washed three times using a washing buffer and incubated with streptavidin-HRP for 30 minutes. Lastly, the membranes were rewashed three times before the substrate was added to develop chemiluminescence conditions. The membrane was then viewed using ChemiDoc XRS (Bio-Rad).