VMAT1

Figure 1. Western blot analysis of SLC18A1 using anti- SLC18A1 antibody. 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: human Hela whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human U-87MG whole cell lysates, Lane 4: human A431 whole cell lysates, Lane 5: human HL-60 cell lysates, Lane 6: human K562 cell lysates. 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 rabbit anti- SLC18A1 antigen affinity purified polyclonal antibody at 0.5 mug/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 SLC18A1 at approximately 56KD. The expected band size for SLC18A1 is at 56KD.

Figure 2. IHC analysis of SLC18A1 using anti-SLC18A1 antibody. SLC18A1 was detected in paraffin-embedded section of human pancreatic 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 1μg/ml rabbit anti-SLC18A1 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.

Figure 3. IHC analysis of SLC18A1 using anti-SLC18A1 Antibody. SLC18A1 was detected in paraffin-embedded section of human rectal 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 1mug/ml rabbit anti-SLC18A1 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.

Figure 4. Flow Cytometry analysis of HL-60 cells using anti-SLC18A1 antibody Overlay histogram showing HL-60 cells stained with (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- SLC18A1 Antibody 1mug/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1mug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.