Anti-ATG14LAntibody(原货号PB0503)

[list_product_images]Figure 1. Western blot analysis of ATG14L using anti-ATG14L antibody (PB9481). 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: 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-ATG14L antigen affinity purified polyclonal antibody (Catalog # PB9481) 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 ATG14L at approximately 59KD. The expected band size for ATG14L is at 59KD.|Figure 2. IHC analysis of ATG14L using anti-ATG14L antibody (PB9481).ATG14L was detected in paraffin-embedded section of Rat Spleen 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-ATG14L Antibody (PB9481) 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 ATG14L using anti-ATG14L antibody (PB9481).ATG14L 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-ATG14L Antibody (PB9481) 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 4. IF analysis of ATG14 using anti-ATG14 antibody (PB9481).
ATG14 was detected in an immunocytochemical section of U2OS cells. Cy3-conjugated Anti-rabbit IgG Secondary Antibody (red)(Catalog#BA1032) was used as secondary antibody. The section was counterstained with DAPI (Catalog # AR1176) (Blue). |Figure 5. IF analysis of ATG14 using anti-ATG14 antibody (PB9481).
ATG14 was detected in a paraffin-embedded section of human lung cancer tissue. Cy3-conjugated Anti-rabbit IgG Secondary Antibody (red)(Catalog#BA1032) was used as secondary antibody. The section was counterstained with DAPI (Catalog # AR1176) (Blue). |Figure 6. IF analysis of ATG14 using anti-ATG14 antibody (PB9481).
ATG14 was detected in a paraffin-embedded section of human colon cancer tissue. Cy3-conjugated Anti-rabbit IgG Secondary Antibody (red)(Catalog#BA1032) was used as secondary antibody. The section was counterstained with DAPI (Catalog # AR1176) (Blue). |Figure 7. IF analysis of ATG14 using anti-ATG14 antibody (PB9481).
ATG14 was detected in a paraffin-embedded section of rat spleen tissue. Cy3-conjugated Anti-rabbit IgG Secondary Antibody (red)(Catalog#BA1032) was used as secondary antibody. The section was counterstained with DAPI (Catalog # AR1176) (Blue). |Figure 8. Flow Cytometry analysis of SiHa cells using anti-ATG14L antibody (PB9481).Overlay histogram showing SiHa cells stained with PB9481 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG14L Antibody (PB9481,1μg/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 9. Flow Cytometry analysis of A431 cells using anti-ATG14L antibody (PB9481).Overlay histogram showing A431 cells stained with PB9481 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG14L Antibody (PB9481,1μg/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 10. Flow Cytometry analysis of PC-3 cells using anti-ATG14L antibody (PB9481).Overlay histogram showing PC-3 cells stained with PB9481 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG14L Antibody (PB9481,1μg/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.[/list_product_images]