Anti-Aspartate beta hydroxylas

[list_product_images]Figure 1. Western blot analysis of ASPH using anti-ASPH antibody (PB0502). 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: Rat Brain Tissue Lysate,Lane 2: Rat Liver Tissue Lysate,Lane 3: HELA Whole Cell Lysate,Lane 4: HEPG2 Whole Cell Lysate,Lane 5: HEPA Whole Cell Lysate. 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-ASPH antigen affinity purified polyclonal antibody (Catalog # PB0502) 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 1:10000 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 ASPH at approximately 100KD. The expected band size for ASPH is at 86KD.|Figure 2. IHC analysis of ASPH using anti-ASPH antibody (PB0502).ASPH was detected in paraffin-embedded section of Human Mammary Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ASPH Antibody (PB0502) 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)(Catalog # SA1022) with DAB as the chromogen. |Figure 3. IHC analysis of ASPH using anti- ASPH antibody (PB0502).
ASPH was detected in immunocytochemical section of A549 cell. 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 1μg/ml rabbit anti- ASPH Antibody ( PB0502) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 4. Flow Cytometry analysis of HeLa cells using anti- ASPH antibody (PB0502).
Overlay histogram showing HeLa cells stained with PB0502 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- ASPH Antibody (PB0502,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.|Figure 5. Flow Cytometry analysis of U87 cells using anti- ASPH antibody (PB0502).
Overlay histogram showing U87 cells stained with PB0502 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti- ASPH Antibody (PB0502,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.[/list_product_images]