Anti-Grainyhead-like protein 1/GRHL1 Antibody Picoband®

Western blot analysis of Grainyhead-like protein 1/GRHL1 using anti-Grainyhead-like protein 1/GRHL1 antibody (A07368-1).
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 T-47D whole cell lysates,
Lane 2: human MDA-MB-453 whole cell lysates,
Lane 3: human MCF-7 whole 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-Grainyhead-like protein 1/GRHL1 antigen affinity purified polyclonal antibody (Catalog # A07368-1) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Grainyhead-like protein 1/GRHL1 at approximately 70KD. The expected band size for Grainyhead-like protein 1/GRHL1 is at 70KD.

IF analysis of Grainyhead-like protein 1/GRHL1 using anti-Grainyhead-like protein 1/GRHL1 antibody (A07368-1).
Grainyhead-like protein 1/GRHL1 was detected in immunocytochemical section of MCF-7 cells. 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 5μg/mL rabbit anti-Grainyhead-like protein 1/GRHL1 Antibody (A07368-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of U251 cells using anti-Grainyhead-like protein 1/GRHL1 antibody (A07368-1).
Overlay histogram showing U251 cells stained with A07368-1 (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-Grainyhead-like protein 1/GRHL1 Antibody (A07368-1, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.