Elisa picokine kits

ELISA kits were used to assay biomarkers in the urine of healthy, OW/OB and post-intervening OW/OB adolescents. ELISA PicoKine™ kits (BosterBio, Pleasanton, CA, USA) were used to detect tumor necrosis factor-α (TNFα) (Cat No. SKU: EK0525), interleukin-6 (IL-6) (Cat No. SKU: EK0410), and endothelin-1 (ET-1) (Cat No. SKU: EK0945) as per manufacturer protocol. Here, 100 ul of standard (# 1—1000 pg/mL, #2—500 pg/mL, #3—250 pg/mL, #4—125 pg/mL, #5—62.5 pg/mL, #6—31.25 pg/mL, #7—15.625 pg/m) and blank control (sample diluent, i.e., 0 pg/mL) were used in triplicate for the generation of the standard curve. We averaged the triplicate readings for each standard, sample, and control, followed by subtracting the average blank control OD reading. Finally, we generated the standard curve and derived the y = mx + c equation to measure the sample’s protein concentration. If data were outside of the detection limit, they were managed by nonparametric methods [16 (link)].

ELISA kits were used to assay biomarkers in the urine of healthy, OW/OB and post-intervening OW/OB adolescents. ELISA PicoKine™ kits (BosterBio, Pleasanton, CA, USA) were used to detect tumor necrosis factor-α (TNFα) (Cat No. SKU: EK0525), interleukin-6 (IL-6) (Cat No. SKU: EK0410), and endothelin-1 (ET-1) (Cat No. SKU: EK0945) as per manufacturer protocol. Here, 100 ul of standard (# 1—1000 pg/mL, #2—500 pg/mL, #3—250 pg/mL, #4—125 pg/mL, #5—62.5 pg/mL, #6—31.25 pg/mL, #7—15.625 pg/m) and blank control (sample diluent, i.e., 0 pg/mL) were used in triplicate for the generation of the standard curve. We averaged the triplicate readings for each standard, sample, and control, followed by subtracting the average blank control OD reading. Finally, we generated the standard curve and derived the y = mx + c equation to measure the sample’s protein concentration. If data were outside of the detection limit, they were managed by nonparametric methods [16 (link)].