High sensitive enzyme linked immunosorbent assay kit

The commercial rat ELISA kit (R&D Systems, USA) was used to evaluate TNF-α, and for IL-6 the Cloud-Clone Corp., High Sensitive Enzyme-linked Immunosorbent Assay Kit (HEA079Ra, USA) was used.
The procedures were performed according to the manufacturer’s instructions.
Fifty mg portions of kidney tissue were cut into small pieces and homogenized in 2 mL of ice-cold PBS with protease inhibitors. The resulting suspension was subjected to two freeze–thaw cycles to further break cell membranes. The homogenates were centrifuged for 5 min at 5000 ×g in 4 °C. After centrifuging, the supernatant was separated and applied to determine TNF-α and IL-6.

The commercial rat ELISA kit (R&D Systems, Minneapolis, MN, USA) was used to evaluate TNF-α whereas for IL-6, the Cloud-Clone Corp. (Houston, TX, USA) High Sensitive Enzyme-linked Immunosorbent Assay Kit (HEA079Ra, Houston, TX, USA) was used.
These markers were measured according to the manufacturer’s instructions. TNF-α and IL-6 concentrations were determined from the standard curves and expressed in pg/mg protein.

High sensitive Enzyme-Linked Immunosorbent Assay kit (Cloud-Clone Corp., Katy, TX USA) was used for in vitro quantitative measurement of human VEGFA in culture medium of HGC-27, AGS, NCI-N87 [N87] and KATO III cells treated as previously described, according to the instruction manual.