Human cxcl1 gro α duoset

For the assay, CCD-1070Sk cells (10⁴) were seeded to 24 well plate. After 24 h, 20 ng/mL IL-17A cytokine alone or in the presence of His₆-ARS-TolA-AVI clones in different concentrations (100, 50, 5, 0.5 ng/mL) were added. After 6, 15, 24, 30, and 48 h, supernatants were collected and levels of Gro-α and IL-6 were measured by Human CXCL1/GRO α DuoSet (R&D Systems, Minneapolis, MN, USA).

Paired plasma samples before the start of therapy (pretreatment) and after ± 4 weeks (median 29 days) of treatment (on‐treatment) were obtained from 9 metastatic breast cancer patients who received palbociclib treatment and 10 metastatic breast cancer patients who received paclitaxel treatment. Written informed consent was obtained from all patients. Mean age was 55.9 years (SD 10.9) for the palbociclib group and 60.1 years (SD 7.4) for the paclitaxel group. The samples were collected in cell stabilizing blood collection tubes (CellSave or Streck Cell‐Free DNA BCT tubes). After collection, the blood was centrifuged twice (1700 g for 10 min and 12,000 g for 10 min). Afterward, the plasma was pooled and directly stored in 2‐ml tubes in −80°C until analyzed. For ELISA, human CXCL1/GROα duo‐set or CCL2 duo‐set or CCL5 duo‐set or MMP1 duo‐set or IGFBP3 duo‐set (R&D Systems) was used to detect the concentrations of CXCL1 or CCL2 or CCL5 or MMP1 or IGFBP3 in the plasma following manufacturer’s instructions.