[list_product_images]Figure 1. Western blot analysis of CEACAM5/Cd66e using anti- CEACAM5/Cd66e antibody (BM0024).
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 # BM0024) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. |Figure 1 For the blank IHC analysis using PBS was detected in paraffin-embedded section of human colon cancer tissue ,Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen. Figure 2 Analysis of CEACAM5/Cd66e. IHC using anti-CEACAM5/Cd66e antibody (BM0024) 1:200 was detected in paraffin-embedded section of human colon cancer tissue. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen. Figure 3 negative control IHC analysis of CEACAM5/Cd66e antibody (BM0024)1:200 was detected in paraffin-embedded section of human placenta tissue. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.[/list_product_images]