Western blot (WB): | 1:500-2000 |
Immunohistochemistry (IHC): | 1:50-400 |
Flow Cytometry (Fixed): | 1:50-200 |
(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 anti- ACTN3 antibody (PB10026). The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: HT1080 whole cell lysates,
Lane 2: rat skeletal muscle tissue lysates,
Lane 3: rat skeletal muscle tissue lysates,
Lane 4: mouse skeletal muscle tissue lysates,
Lane 5: mouse skeletal muscle tissue lysates.
Use rabbit anti- ACTN3 1:1000, probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002). 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/Alpha Actinin 3 using anti- ACTN3/Alpha Actinin 3 antibody (PB10026).ACTN3/Alpha Actinin 3 was detected in paraffin-embedded section of mouse skeletal muscle 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- ACTN3/Alpha Actinin 3 Antibody (PB10026) 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.
Figure 3. IHC analysis of ACTN3/Alpha Actinin 3 using anti- ACTN3/Alpha Actinin 3 antibody (PB10026).ACTN3/Alpha Actinin 3 was detected in paraffin-embedded section of rat skeletal muscle 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- ACTN3/Alpha Actinin 3 Antibody (PB10026) 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.
Figure 4. IHC analysis of ACTN3/Alpha Actinin 3 using anti- ACTN3/Alpha Actinin 3 antibody (PB10026).ACTN3/Alpha Actinin 3 was detected in paraffin-embedded section of human lung cancer 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- ACTN3/Alpha Actinin 3 Antibody (PB10026) 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.
Figure 5. Flow Cytometry analysis of WISH cells using anti-ACTN3 antibody (PB10026).Overlay histogram showing WISH cells stained with PB10026 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ACTN3 Antibody (PB10026, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 1. Western blot analysis of anti- ACTN3 antibody (PB10026). The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: HT1080 whole cell lysates,
Lane 2: rat skeletal muscle tissue lysates,
Lane 3: rat skeletal muscle tissue lysates,
Lane 4: mouse skeletal muscle tissue lysates,
Lane 5: mouse skeletal muscle tissue lysates.
Use rabbit anti- ACTN3 1:1000, probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002). 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/Alpha Actinin 3 using anti- ACTN3/Alpha Actinin 3 antibody (PB10026).ACTN3/Alpha Actinin 3 was detected in paraffin-embedded section of mouse skeletal muscle 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- ACTN3/Alpha Actinin 3 Antibody (PB10026) 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.
Figure 3. IHC analysis of ACTN3/Alpha Actinin 3 using anti- ACTN3/Alpha Actinin 3 antibody (PB10026).ACTN3/Alpha Actinin 3 was detected in paraffin-embedded section of rat skeletal muscle 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- ACTN3/Alpha Actinin 3 Antibody (PB10026) 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.
Figure 4. IHC analysis of ACTN3/Alpha Actinin 3 using anti- ACTN3/Alpha Actinin 3 antibody (PB10026).ACTN3/Alpha Actinin 3 was detected in paraffin-embedded section of human lung cancer 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- ACTN3/Alpha Actinin 3 Antibody (PB10026) 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.
Figure 5. Flow Cytometry analysis of WISH cells using anti-ACTN3 antibody (PB10026).Overlay histogram showing WISH cells stained with PB10026 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ACTN3 Antibody (PB10026, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.