Western blot analysis of EHD1 using anti-EHD1 antibody (A02168-2). 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 T-47D whole cell lysates, Lane 2: human Caco-2 whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: monkey COS-7 whole cell lysates, Lane 6: mouse testis tissue lysates, Lane 7: mouse kidney tissue lysates, Lane 8: mouse lung tissue lysates, Lane 9: mouse ovary 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 rabbit anti-EHD1 antigen affinity purified polyclonal antibody (Catalog # A02168-2) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for EHD1 at approximately 61 kDa. The expected band size for EHD1 is at 61 kDa.

IF analysis of EHD1 using anti-EHD1 antibody (A02168-2). EHD1 was detected in an immunocytochemical section of SiHa 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 rabbit anti-EHD1 Antibody (A02168-2) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.