Assay principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Revvity low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.

Intra assay (n=24)

Inter assay (n=4)

CCL5 secretion in PBMCs stimulated with LPS, IL1ß and TNFa

PBMCs were plated at 100,000 cells/well in a 96 well plate, and stimulated for 24h with LPS, IL1ß and TNFa respectively used at 200 µg/mL, 10 ng/mL and 10 ng/mL. Supernatants were diluted to 1/3, and 16 µL of the diluted samples were transferred into a 384 well (SV) white detection plate to be analyzed by with the Human CCL5 Assay.

CCL5 secretion in LX2 stimulated with TGFß

LX2 cells were plated at 400,000 cells/well in a 6 well plate, cultured for 3 days, then starved for 1 day before 48 hours of stimulation with TGFß at 0.5ng/ml. Next, 16 µL of supernatant were transferred into a 384 well (SV) white detection plate to be analyzed with the Human CCL5 Assay.