Anti-HDAC4 Antibody Picoband®

Western blot analysis of HDAC4 using anti-HDAC4 antibody (A00971-1).
Electrophoresis was performed on a 8% 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 Jurkat whole cell lysates,
Lane 2: human THP-1 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human SH-SY5Y whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse NIH3T3 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-HDAC4 antigen affinity purified polyclonal antibody (A00971-1) 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 HDAC4 at approximately 140 kDa. The expected band size for HDAC4 is at 119 kDa.

IHC analysis of HDAC4 using anti-HDAC4 antibody (A00971-1).
HDAC4 was detected in a paraffin-embedded section of human cerebral cortex tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HDAC4 Antibody (A00971-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of HDAC4 using anti-HDAC4 antibody (A00971-1) and anti-Alpha Tubulin antibody (M03989-3).
HDAC4 was detected in an immunocytochemical section of Hela 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 5 μg/mL rabbit anti-HDAC4 Antibody (A00971-1) and mouse anti-Alpha Tubulin antibody (M03989-3) overnight at 4°C. Fluoro488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating HDAC4 in Jurkat whole cell lysate.
Western blot analysis of HDAC4 using anti-HDAC4 antibody (A00971-1).
Lane 1: Jurkat whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-HDAC4 antibody in Jurkat whole cell lysate,
Lane 3: anti-HDAC4 antibody (2μg) + Jurkat whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-HDAC4 antigen affinity purified polyclonal antibody (A00971-1) 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 HDAC4 at approximately 140 kDa. The expected band size for HDAC4 is at 119 kDa.