25 ug total protein per lane of various lysates (see on figure) probed with CCR7 monoclonal antibody, unconjugated (bsm-52774R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CCR7 Monoclonal Antibody, Unconjugated(bsm-52774R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CCR7 Monoclonal Antibody, Unconjugated(bsm-52774R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CCR7 Monoclonal Antibody, Unconjugated(bsm-52774R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.

ICC staining of CCR7 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (bsm-52774R, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Blank control（black line）:Jurkat. Primary Antibody (green line): Mouse Anti-CCR7 antibody (bsm-52774R) Dilution:1ug/Test; Secondary Antibody（white blue line）: Goat anti-Mouse IgG-AF488 Dilution: 0.5ug/Test. Isotype control（orange line）: Normal Rabbit IgG Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.