Anti-PD-ECGF/TYMP Antibody Picoband®

Western blot analysis of PD-ECGF/TYMP using anti-PD-ECGF/TYMP antibody (A06329-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A431 whole cell lysates,
Lane 2: human SiHa whole cell lysates,
Lane 3: human THP-1 whole cell lysates.
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-PD-ECGF/TYMP antigen affinity purified polyclonal antibody (A06329-1) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for PD-ECGF/TYMP at approximately 50 kDa. The expected band size for PD-ECGF/TYMP is at 50 kDa.

Immunoprecipitating PD-ECGF/TYMP in SiHa whole cell lysate.
Western blot analysis of PD-ECGF/TYMP using anti-PD-ECGF/TYMP antibody (A06329-1).
Lane 1: SiHa whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-PD-ECGF/TYMP antibody in SiHa whole cell lysate,
Lane 3: anti-PD-ECGF/TYMP antibody (2μg) + SiHa whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PD-ECGF/TYMP antigen affinity purified polyclonal antibody (A06329-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for PD-ECGF/TYMP at approximately 50 kDa. The expected band size for PD-ECGF/TYMP is at 50 kDa.

Flow Cytometry analysis of A431 cells using anti-PD-ECGF/TYMP antibody (A06329-1).
Overlay histogram showing A431 cells stained with A06329-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PD-ECGF/TYMP Antibody (A06329-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of SiHa cells using anti-PD-ECGF/TYMP antibody (A06329-1).
Overlay histogram showing SiHa cells stained with A06329-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PD-ECGF/TYMP Antibody (A06329-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.