Anti-MIEF1 Antibody Picoband®

Western blot analysis of MIEF1 using anti-MIEF1 antibody (A06887-1).
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 30ug of sample under reducing conditions.
Lane 1: human HT1080 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human HL-60 whole cell lysates,
Lane 5: human HEK293 whole cell lysates,
Lane 6: human U-937 whole cell lysates,
Lane 7: human HepG2 whole cell lysates,
Lane 8: rat heart tissue lysates,
Lane 9: rat kidney tissue lysates,
Lane 10: mouse kidney tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-MIEF1 antigen affinity purified polyclonal antibody (Catalog # A06887-1) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MIEF1 at approximately 51KD. The expected band size for MIEF1 is at 51KD.

IHC analysis of MIEF1 using anti-MIEF1 antibody (A06887-1).
MIEF1 was detected in paraffin-embedded section of human renal clear cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-MIEF1 Antibody (A06887-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of MIEF1 using anti-MIEF1 antibody (A06887-1).
MIEF1 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-MIEF1 Antibody (A06887-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IF analysis of MIEF1 using anti-MIEF1 antibody (A06887-1).
MIEF1 was detected in immunocytochemical section of U20S 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-MIEF1 Antibody (A06887-1) 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.

Flow Cytometry analysis of SiHa cells using anti-MIEF1 antibody (A06887-1).
Overlay histogram showing SiHa cells stained with A06887-1 (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-MIEF1 Antibody (A06887-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Immunoprecipitating (IP) MIEF1 in K562 whole cell lysate.
Western blot analysis of MIEF1 using anti-MIEF1 antibody (A06887-1);
Lane 1: K562 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-MIEF1 antibody in K562 whole cell lysate;
Lane 3: anti-MIEF1 antibody (2μg) + K562 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-MIEF1 antigen affinity purified polyclonal antibody (A06887-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Light Chain). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for MIEF1 at approximately 51 kDa. The expected band size for MIEF1 is at 51 kDa.