Application
Fig1: Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody at 1/5,000 dilution.
Lane 1: A549 cell lysate,
Lane 2: MDA-MB-231 cell lysate,
Lane 3: SK-Br-3 cell lysate (low expression),
Lane 4: Mouse brain tissue lysate,
Lane 5: Mouse heart tissue lysate,
Lane 6: Rat brain tissue lysate,
Lane 7: Rat heart tissue lysate,
Lysates/proteins at 20 µg/Lane.
Exposure time: Lane 1-3: 1 minute 21 seconds; Lane 4-7: 43 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
Application
Fig2: ICC staining Neuropilin-1 in SHG-44 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig3: ICC staining Neuropilin-1 in MCF-7 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig4: ICC staining Neuropilin-1 in HUVEC cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Neuropilin 1 antibody at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour 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.
Application
Fig6:Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Neuropilin 1 antibody at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour 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.