Anti-Lipoamide Dehydrogenase/DLD Antibody Picoband®

Anti-DLD antibody, PA1463, IHC(P)
IHC(P): Human Mammary Cancer Tissue

Anti-Lipoamide Dehydrogenase antibody, PA1463, ICC
ICC: MCF-7 Cell

Western blot analysis of DLD using anti-DLD antibody (PA1463).
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 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human SiHa whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: 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-DLD antigen affinity purified polyclonal antibody (Catalog # PA1463) 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 DLD at approximately 54 kDa. The expected band size for DLD is at 54 kDa.

IF analysis of DLD using anti-DLD antibody (PA1463)
DLD was detected in paraffin-embedded section of rat liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-DLD Antibody (PA1463) 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.

IF analysis of DLD using anti-DLD antibody (PA1463).
DLD was detected in immunocytochemical section of HCT116 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-DLD Antibody (PA1463) 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.

Western blot analysis of DLD using anti-DLD antibody (PA1463).
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 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human SiHa whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: 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-DLD antigen affinity purified polyclonal antibody (PA1463) 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-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for DLD at approximately 54 kDa. The expected band size for DLD is at 54 kDa.