Western blot (WB): | 1:500-2000 |
Immunohistochemistry (IHC): | 1:50-400 |
Immunocytochemistry/Immunofluorescence (ICC/IF): | 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 CTNNA1 using anti-CTNNA1 antibody (PB9137). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: Recombinant Human CTNNA1 Protein 0.5ng. 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 rabbit anti-CTNNA1 antigen affinity purified polyclonal antibody (Catalog # PB9137) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CTNNA1 at approximately 47KD. The expected band size for CTNNA1 is at 47KD.
Figure 2. Western blot analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137). Electrophoresis was performed on a 5-20% 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 Liver Tissue LysateLane 2: Rat Lung Tissue LysateLane 3: Rat Cardiac Muscle Tissue LysateLane 4: NIH3T3 Whole Cell LysateLane 5: PC-12 Whole Cell LysateLane 6: HEPG2 Whole Cell LysateLane 7: HELA Whole Cell LysateLane 8: MCF-7 Whole Cell LysateLane 9: HEPA Whole Cell LysateAfter 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 rabbit anti-CTNNA1 antigen affinity purified polyclonal antibody (Catalog # PB9137) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CTNNA1 at approximately 100KD. The expected band size for CTNNA1 is at 100KD.
Figure 3. IHC analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137).CTNNA1 was detected in paraffin-embedded section of rat intestine 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 rabbit anti-CTNNA1 Antibody (PB9137) 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 CTNNA1 using anti-CTNNA1 antibody (PB9137).CTNNA1 was detected in paraffin-embedded section of human mammary 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 rabbit anti-CTNNA1 Antibody (PB9137) 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. IHC analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137).CTNNA1 was detected in paraffin-embedded section of mouse intestine 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 rabbit anti-CTNNA1 Antibody (PB9137) 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 6. Flow Cytometry analysis of U-87 cells using anti-CTNNA1 antibody (PB9137).Overlay histogram showing U-87 cells stained with PB9137 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CTNNA1 Antibody (PB9137, 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 7. ICC analysis using anti- CTNNA1 antibody (PB9137) was detected in immersion fixed U2OS cell line . Cells were stained using the Dylight488-conjugated Anti-rabbit IgG Secondary Antibody (green)(Catalog # BA1127) and counterstained with DAPI (blue).
Figure 1. Western blot analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: Recombinant Human CTNNA1 Protein 0.5ng. 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 rabbit anti-CTNNA1 antigen affinity purified polyclonal antibody (Catalog # PB9137) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CTNNA1 at approximately 47KD. The expected band size for CTNNA1 is at 47KD.
Figure 2. Western blot analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137). Electrophoresis was performed on a 5-20% 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 Liver Tissue LysateLane 2: Rat Lung Tissue LysateLane 3: Rat Cardiac Muscle Tissue LysateLane 4: NIH3T3 Whole Cell LysateLane 5: PC-12 Whole Cell LysateLane 6: HEPG2 Whole Cell LysateLane 7: HELA Whole Cell LysateLane 8: MCF-7 Whole Cell LysateLane 9: HEPA Whole Cell LysateAfter 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 rabbit anti-CTNNA1 antigen affinity purified polyclonal antibody (Catalog # PB9137) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CTNNA1 at approximately 100KD. The expected band size for CTNNA1 is at 100KD.
Figure 3. IHC analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137).CTNNA1 was detected in paraffin-embedded section of rat intestine 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 rabbit anti-CTNNA1 Antibody (PB9137) 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 CTNNA1 using anti-CTNNA1 antibody (PB9137).CTNNA1 was detected in paraffin-embedded section of human mammary 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 rabbit anti-CTNNA1 Antibody (PB9137) 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. IHC analysis of CTNNA1 using anti-CTNNA1 antibody (PB9137).CTNNA1 was detected in paraffin-embedded section of mouse intestine 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 rabbit anti-CTNNA1 Antibody (PB9137) 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 6. Flow Cytometry analysis of U-87 cells using anti-CTNNA1 antibody (PB9137).Overlay histogram showing U-87 cells stained with PB9137 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CTNNA1 Antibody (PB9137, 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 7. ICC analysis using anti- CTNNA1 antibody (PB9137) was detected in immersion fixed U2OS cell line . Cells were stained using the Dylight488-conjugated Anti-rabbit IgG Secondary Antibody (green)(Catalog # BA1127) and counterstained with DAPI (blue).