Anti-PDH E1 Alpha/PDHA1 Antibody

Western blot analysis of PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
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 293T whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse kidney 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-PDH E1 Alpha/PDHA1 antigen affinity purified polyclonal antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 at approximately 40 kDa. The expected band size for PDH E1 Alpha/PDHA1 is at 43 kDa.

IHC analysis of PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in a paraffin-embedded section of human ovarian serous adenocarcinoma 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 1:100 rabbit anti-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in a paraffin-embedded section of human breast 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 1:100 rabbit anti-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in a paraffin-embedded section of human spleen 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 1:100 rabbit anti-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in a paraffin-embedded section of human testicular germ cell tumors 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 1:100 rabbit anti-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in a paraffin-embedded section of human colorectal adenocarcinoma 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 1:100 rabbit anti-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in a paraffin-embedded section of human appendix carcinoid 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 1:100 rabbit anti-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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 PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4).
PDH E1 Alpha/PDHA1 was detected in an immunocytochemical section of chuman Hela ells. 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-PDH E1 Alpha/PDHA1 Antibody (A01906-4) 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.

Immunoprecipitating (IP) PDH E1 Alpha/PDHA1 in HepG2 whole cell lysate.
Western blot analysis of PDH E1 Alpha/PDHA1 using anti-PDH E1 Alpha/PDHA1 antibody (A01906-4);
Lane 1: HepG2 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-PDH E1 Alpha/PDHA1 antibody in HepG2 whole cell lysate;
Lane 3: anti-PDH E1 Alpha/PDHA1 antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PDH E1 Alpha/PDHA1 antigen affinity purified polyclonal antibody (A01906-4) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Light Chiain). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for PDH E1 Alpha/PDHA1 at approximately 40 kDa. The expected band size for PDH E1 Alpha/PDHA1 is at 43 kDa.