Anti-Ataxin 3/ATXN3 Antibody(原

[list_product_images]Figure 1. Western blot analysis of Ataxin 3 using anti-Ataxin 3 antibody (PB9423). 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 LysateLane 2: COLO320 Whole Cell LysateLane 3: HELA 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-Ataxin 3 antigen affinity purified polyclonal antibody (Catalog # PB9423) 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 Ataxin 3 at approximately 42KD. The expected band size for Ataxin 3 is at 42KD.|Figure 2. IHC analysis of Ataxin 3 using anti-Ataxin 3 antibody (PB9423).Ataxin 3 was detected in paraffin-embedded section of Human Lung 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-Ataxin 3 Antibody (PB9423) 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. Flow Cytometry analysis of A549 cells using anti-ATXN3 antibody (PB9423).Overlay histogram showing A549 cells stained with PB9423 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATXN3 Antibody (PB9423,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 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 4. Flow Cytometry analysis of SiHa cells using anti-ATXN3 antibody (PB9423).Overlay histogram showing SiHa cells stained with PB9423 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATXN3 Antibody (PB9423,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 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. IF analysis of ATXN3 using anti- ATXN3 antibody (PB9423).
ATXN3 was detected in immunocytochemical section of MCF-7 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 2μg/mL rabbit anti- ATXN3 Antibody (PB9423) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.[/list_product_images]