NAA60 Antibody / N-alpha-acetyltransferase 60 (FY13134)

Immunofluorescent staining of NAA60 using anti-NAA60 antibody (green). NAA60 was detected in an immunocytochemical section of cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-NAA60 antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of NAA60 using anti-NAA60 antibody. Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human MCF-7 whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human HEL whole cell lysates, Lane 4: human Caco-2 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat RH35 whole cell lysates, Lane 7: mouse brain 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-NAA60 antibody at 0.5 ug/ml overnight at 4oC, 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 was developed using an ECL Plus Western Blotting Substrate. The expected molecular weight of NAA60 is ~27 kDa.

Immunohistochemical staining of NAA60 using anti-NAA60 antibody. NAA60 was detected in a paraffin-embedded section of human prostate 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 ug/ml rabbit anti-NAA60 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Immunoprecipitating NAA60 in MCF-7 whole cell lysate. Western blot analysis of NAA60 using anti-NAA60 antibody. Lane 1: MCF-7 whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-NAA60 antibody in MCF-7 whole cell lysate, Lane 3: anti-NAA60 antibody (2ug) + MCF-7 whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-NAA60 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. The expected molecular weight of NAA60 is at 27 kDa.