Anti-FOXJ2 Antibody Picoband®

Western blot analysis of FOXJ2 using anti-FOXJ2 antibody (A06789-1).
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 U2OS whole cell lysates,
Lane 2: human SIHA whole cell lysates,
Lane 3: human U-87MG whole cell lysates,
Lane 4: rat heart tissue lysates,
Lane 5: rat lung tissue lysates,
Lane 6: mouse heart tissue 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-FOXJ2 antigen affinity purified polyclonal antibody (A06789-1) 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 FOXJ2 at approximately 70 kDa. The expected band size for FOXJ2 is at 62 kDa.

IHC analysis of FOXJ2 using anti-FOXJ2 antibody (A06789-1).
FOXJ2 was detected in a paraffin-embedded section of rat brain 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-FOXJ2 Antibody (A06789-1) 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.

IHC analysis of FOXJ2 using anti-FOXJ2 antibody (A06789-1).
FOXJ2 was detected in a paraffin-embedded section of human brain 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-FOXJ2 Antibody (A06789-1) 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.

IHC analysis of FOXJ2 using anti-FOXJ2 antibody (A06789-1).
FOXJ2 was detected in a paraffin-embedded section of human testis 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-FOXJ2 Antibody (A06789-1) 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 FOXJ2 using anti-FOXJ2 antibody (A06789-1) and anti-Alpha Tubulin antibody (M03989-3).
FOXJ2 was detected in an immunocytochemical section of U2OS 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-FOXJ2 Antibody (A06789-1) and mouse anti-Alpha Tubulin antibody (M03989-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and Fluoro488 Conjugated Goat Anti-Mouse IgG (BA1126) 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 FOXJ2 in U2OS whole cell lysate.
Western blot analysis of FOXJ2 using anti-FOXJ2 antibody (A06789-1).
Lane 1: U2OS whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-FOXJ2 antibody in U2OS whole cell lysate,
Lane 3: anti-FOXJ2 antibody (2μg) + U2OS whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-FOXJ2 antigen affinity purified polyclonal antibody (A06789-1) 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 FOXJ2 at approximately 70 kDa. The expected band size for FOXJ2 is at 62 kDa.

Flow Cytometry analysis of MCF-7 cells using anti-FOXJ2 antibody (A06789-1).
Overlay histogram showing MCF-7 cells stained with A06789-1 (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-FOXJ2 Antibody (A06789-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. Fluoro488 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.