Figure 1. Western blot analysis of CEACAM5/Cd66e using anti-CEACAM5/Cd66e antibody (MA1023). 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 50ug of sample under reducing conditions. Lane 1: Recombinant Human CEACAM5/Cd66e Protein 10ng Lane 2; Recombinant Human CEACAM5/Cd66e Protein 5ng Lane 3: Recombinant Human CEACAM5/Cd66e Protein 2.5ng Lane 4: Recombinant Human CEACAM5/Cd66e Protein 1.25ng After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-CEACAM5/Cd66e antigen affinity purified monoclonal antibody (Catalog # MA1023) at 0.5 microg/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system.