Western blot analysis of Neuropilin 1 using anti-Neuropilin 1 antibody (M01324-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human placenta tissue lysates, Lane 2: human U-87MG whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat stomach tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Neuropilin 1 antigen affinity purified monoclonal antibody (Catalog # M01324-2) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Neuropilin 1 at approximately 120 kDa. The expected band size for Neuropilin 1 is at 120 kDa.

IF analysis of Neuropilin 1 using anti-Neuropilin 1 antibody (M01324-2). Neuropilin 1 was detected in an immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-Neuropilin 1 Antibody (M01324-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.