Sample:
Lane 1: Thymus (Mouse) Lysate at 40 ug
Lane 2: Spleen (Mouse) Lysate at 40 ug
Lane 3: Lymph node (Mouse) Lysate at 40 ug
Lane 4: Lung (Mouse) Lysate at 40 ug
Lane 5: Thymus (Rat) Lysate at 40 ug
Lane 6: Spleen (Rat) Lysate at 40 ug
Lane 7: Lymph node (Rat) Lysate at 40 ug
Lane 8: Lung (Rat) Lysate at 40 ug
Lane 9: Jurkat (Human) Cell Lysate at 30 ug
Lane 10: MCF-7 (Human) Cell Lysate at 30 ug
Lane 11: Hela (Human) Cell Lysate at 30 ug
Lane 12: HL60 (Human) Cell Lysate at 30 ug
Primary: Anti-CCR7 (bsm-52774R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD

Western blot analysis of CCR7 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (bsm-52774R, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Jurkat cell lysate Lane 2: MCF-7 cell lysate Lane 3: PC-12 cell lysate Lane 4: Daudi cell lysate

All lanes: Western blot analysis of CCR7 with anti-CCR7 antibody at 1:5,000 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (10 µg).
Lane 3/4: CCR7 fragment 1 knockdown Hela whole cell lysate (10 µg).
Lane 5/6: CCR7 fragment 2 knockdown Hela whole cell lysate (10 µg).

Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CCR7 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52774R, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CCR7 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody ( bsm-52774R, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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).

ICC staining of CCR7 in MCF-7 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.