Rab5/RAB5A Rabbit Polyclonal Antibody

IHC analysis of Rab5 using anti-Rab5 antibody. Rab5 was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Rab5 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of Rab5 using anti-Rab5 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: Rat Brain Tissue Lysate at 50 ug, Lane 2: Mouse Brain Tissue Lysate at 50 ug, Lane 3: HELA Whole Cell Lysate at 40 ug, Lane 4: SW620 Whole Cell Lysate at 40 ug, Lane 5: 293T Whole Cell Lysate at 40 ug, Lane 6: JURKAT Whole Cell Lysate at 40 ug. 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-Rab5 antigen affinity purified polyclonal antibody 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 with Tanon 5200 system. A specific band was detected for Rab5 at approximately 24 kDa. The expected band size for Rab5 is at 24 kDa.