Anti-Alpha Actinin/ACTN1 Antibody Picoband®

Western blot analysis of ACTN1 using anti-ACTN1 antibody (A02635-1).
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 Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: rat heart tissue lysates,
Lane 4: rat spleen tissue lysates,
Lane 5: rat C6 whole cell lysates,
Lane 6: mouse heart tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse NIH/3T3 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-ACTN1 antigen affinity purified polyclonal antibody (A02635-1) 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 ACTN1 at approximately 105 kDa. The expected band size for ACTN1 is at 103 kDa.

Western blot analysis of Alpha Actinin/ACTN1 using anti-Alpha Actinin/ACTN1 antibody (A02635-1).
Electrophoresis was performed on a 8% 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 A431 whole cell lysates,
Lane 2: human SiHa whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: rat small intestine tissue lysates,
Lane 6: rat PC-12 whole cell lysates,
Lane 7: mouse small intestine tissue lysates,
Lane 8: mouse NIH/3T3 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-Alpha Actinin/ACTN1 antigen affinity purified polyclonal antibody (A02635-1) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Alpha Actinin/ACTN1 at approximately 103 kDa. The expected band size for Alpha Actinin/ACTN1 is at 103 kDa.

IHC analysis of Alpha Actinin/ACTN1 using anti-Alpha Actinin/ACTN1 antibody (A02635-1).
Alpha Actinin/ACTN1 was detected in a paraffin-embedded section of human colon 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-Alpha Actinin/ACTN1 Antibody (A02635-1) 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 Alpha Actinin/ACTN1 using anti-Alpha Actinin/ACTN1 antibody (A02635-1).
Alpha Actinin/ACTN1 was detected in a paraffin-embedded section of mouse heart 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-Alpha Actinin/ACTN1 Antibody (A02635-1) 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 Alpha Actinin/ACTN1 using anti-Alpha Actinin/ACTN1 antibody (A02635-1).
Alpha Actinin/ACTN1 was detected in an immunocytochemical section of SIHA 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 5 μg/mL rabbit anti-Alpha Actinin/ACTN1 Antibody (A02635-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) 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.

Flow Cytometry analysis of PC-12 cells using anti-Alpha Actinin/ACTN1 antibody (A02635-1).
Overlay histogram showing PC-12 cells stained with A02635-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Alpha Actinin/ACTN1 Antibody (A02635-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.