Ccr5 antibody

Recombinant CCL11 was coated onto the wells of maxisorp 96-well microtiter plates and incubated for 18 hours at 4°C. Each well was washed three times with PBST (0.2% Tween-20 in PBS). Thereafter, the plates were blocked with 2% BSA in PBST for 2 hours. Cell lysates were added to the plates and incubated for 2 hours. The wells were then washed three times [27 (link)]. Preparations of the CCR5 antibody (Santa Cruz Biotechnology) in blocking solution were added to the plates and allowed to react for 2 hours. After washing, horseradish peroxidase (HRP)-linked antibody (Cell signaling Technology) was added and the lysates were incubated for 2 hours; this was followed by five rounds of washes. The reaction was developed with 100 μL tetramethylbenzidine (TMB) substrate solution and stopped with 100 μL of 1 N H₂SO₄. The absorbance in the microtiter plates was measured at 450 nm using a microplate reader (Infinite 200 PRO; Tecan Life Sciences, Zürich, Switzerland).