Western blot (WB): | 1:500-2000 |
Immunohistochemistry (IHC): | 1:50-400 |
(Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user. |
Figure 1. Western blot analysis of ACTN3 using anti-ACTN3 antibody (M02693). The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat skeletal muscle tissue,
Lane 2: mouse skeletal muscle tissue. 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 mouse anti-ACTN3 antigen affinity purified monoclonal antibody (Catalog # M02693) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system.A specific band was detected for ACTN3 at approximately 103KD. The expected band size for ACTN3 is at 103KD.
Figure 2. IHC analysis of ACTN3 using anti-ACTN3 antibody (M02693).ACTN3 was detected in paraffin-embedded section of human skeletal muscle tissue. 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 mouse anti-ACTN3 Antibody (M02693) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Figure 1. Western blot analysis of ACTN3 using anti-ACTN3 antibody (M02693). The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat skeletal muscle tissue,
Lane 2: mouse skeletal muscle tissue. 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 mouse anti-ACTN3 antigen affinity purified monoclonal antibody (Catalog # M02693) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system.A specific band was detected for ACTN3 at approximately 103KD. The expected band size for ACTN3 is at 103KD.
Figure 2. IHC analysis of ACTN3 using anti-ACTN3 antibody (M02693).ACTN3 was detected in paraffin-embedded section of human skeletal muscle tissue. 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 mouse anti-ACTN3 Antibody (M02693) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.