CCR6/CD196 Polyclonal Antibody

Paraformaldehyde-fixed, paraffin embedded human liver cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with CCR6 Polyclonal Antibody, Unconjugated (bs-1542R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary (bs-0295G-AF594) at [1:500] for 90 minutes and DAPI staining of the nuclei.

HepG2 cells were\u00a0 incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with CCR6 Polyclonal Antibody(bs-1542R)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).

Lane 1: Human HepG2 cell lysates; Lane 2: Human MCF-7 cell lysates probed with CCR6 Polyclonal Antibody, Unconjugated (bs-1542R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.