Human rantes ccl5 elisa kit

2 ×10⁶ cells expressing either shGFP or shNotch1 were seeded in 10 cm plates with 10% FBS DMEM complete media, and cultured for one day. Cells were then washed three times with PBS before addition of serum free media. After three days in culture, supernatants were collected and the volumes were adjusted by total protein amount in cell lysates. Chemokines were quantified as pg/μg total protein in whole cell lysates. C-arrays and ELISA assays were performed according to manufacturer’s instructions. Human Cytokine Array C5, Mouse Cytokine Array C3, Human IL-8 ELISA Kit (ELH-IL8-1), Human RANTES (CCL5) ELISA Kit (ELH-RANTES-1), Mouse RANTES ELISA Kit (ELM-RANTES-1), Human IL-6 ELISA Kit (ELH-IL6-1), Mouse IL-6 ELISA Kit (ELM-IL6-1), Mouse MIP-2 ELISA Kit (ELM-MIP2-1) were all from Raybiotech.

Conditioned medium was collected from mono- and co-cultures of either 2D adherent cells or 3D tumoroids cultures. CCL5 secretion levels were measured using the Human RANTES (CCL5) ELISA kit (RayBiotech) following manufacturer’s instructions.