Anti-Pyruvate Carboxylase/PC Antibody

Western blot analysis of Pyruvate Carboxylase/PC using anti-Pyruvate Carboxylase/PC antibody (A03853-2).
Electrophoresis was performed on a 12% 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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human SiHa whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat RH-35 whole cell lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse Hepa1-6 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-Pyruvate Carboxylase/PC antigen affinity purified polyclonal antibody (A03853-2) at 1:1000 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 Pyruvate Carboxylase/PC at approximately 130 kDa. The expected band size for Pyruvate Carboxylase/PC is at 130 kDa.

IHC analysis of Pyruvate Carboxylase/PC using anti-Pyruvate Carboxylase/PC antibody (A03853-2).
Pyruvate Carboxylase/PC was detected in a paraffin-embedded section of human liver cancer 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-Pyruvate Carboxylase/PC Antibody (A03853-2) 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 Pyruvate Carboxylase/PC using anti-Pyruvate Carboxylase/PC antibody (A03853-2).
Pyruvate Carboxylase/PC was detected in a paraffin-embedded section of human liver 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-Pyruvate Carboxylase/PC Antibody (A03853-2) 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 Pyruvate Carboxylase/PC using anti-Pyruvate Carboxylase/PC antibody (A03853-2).
Pyruvate Carboxylase/PC 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 1:100 rabbit anti-Pyruvate Carboxylase/PC Antibody (A03853-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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.