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. Flow Cytometry analysis of ANA-1 cells using anti-ARSA antibody (M02583).Overlay histogram showing ANA-1 cells stained with M02583 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ARSA Antibody (M02583, 1:100) for 30 min at 20°C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 2. Western blot analysis of ARSA using anti-ARSA antibody (M02583). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat testis tissue lysate,
Lane 2: rat liver tissue lysate,
Lane 3: rat brain tissue lysate,
Lane 4: rat lung tissue lysate,
Lane 5: mouse testis tissue lysate,
Lane 6: mouse liver tissue lysate,
Lane 7: mouse brain tissue lysate,
Lane 8: mouse lung tissue lysate,
Lane 9: mouse HEPA1-6 whole cell lysate. 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-ARSA antigen affinity purified monoclonal antibody (Catalog # M02583) 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.
Figure 3. Western blot analysis of ARSA using anti-ARSA antibody (M02583). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human A375 whole cell lysate,
Lane 2: human A549 whole cell lysate,
Lane 3: human SMMC-7721 whole cell lysate.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-ARSA antigen affinity purified monoclonal antibody (Catalog # M02583) 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.
Figure 4. Flow Cytometry analysis of Raji cells using anti-ARSA antibody (M02583).
Overlay histogram showing Raji cells stained with M02583 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ARSA Antibody (M02583, 1:100) for 30 min at 20°C. DyLight488 conjugated goat anti-mouse IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 5. IHC analysis of ARSA using anti-ARSA antibody (M02583).
ARSA was detected in paraffin-embedded section of human lung cancer 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-ARSA Antibody (M02583) 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 # SA1022) with DAB as the chromogen.
Figure 1. Flow Cytometry analysis of ANA-1 cells using anti-ARSA antibody (M02583).Overlay histogram showing ANA-1 cells stained with M02583 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ARSA Antibody (M02583, 1:100) for 30 min at 20°C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 2. Western blot analysis of ARSA using anti-ARSA antibody (M02583). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat testis tissue lysate,
Lane 2: rat liver tissue lysate,
Lane 3: rat brain tissue lysate,
Lane 4: rat lung tissue lysate,
Lane 5: mouse testis tissue lysate,
Lane 6: mouse liver tissue lysate,
Lane 7: mouse brain tissue lysate,
Lane 8: mouse lung tissue lysate,
Lane 9: mouse HEPA1-6 whole cell lysate. 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-ARSA antigen affinity purified monoclonal antibody (Catalog # M02583) 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.
Figure 3. Western blot analysis of ARSA using anti-ARSA antibody (M02583). Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human A375 whole cell lysate,
Lane 2: human A549 whole cell lysate,
Lane 3: human SMMC-7721 whole cell lysate.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-ARSA antigen affinity purified monoclonal antibody (Catalog # M02583) 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.
Figure 4. Flow Cytometry analysis of Raji cells using anti-ARSA antibody (M02583).
Overlay histogram showing Raji cells stained with M02583 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ARSA Antibody (M02583, 1:100) for 30 min at 20°C. DyLight488 conjugated goat anti-mouse IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 5. IHC analysis of ARSA using anti-ARSA antibody (M02583).
ARSA was detected in paraffin-embedded section of human lung cancer 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-ARSA Antibody (M02583) 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 # SA1022) with DAB as the chromogen.