Anti-CD3 epsilon/CD3E Antibody Picoband™

Figure 1. Western blot analysis of CD3E using anti-CD3E antibody (PB9093).
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 30 ug of sample under reducing conditions.
Lane 1: human MOLT-4 whole cell lysates,
Lane 2: human Jurkat 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-CD3E antigen affinity purified polyclonal antibody (Catalog # PB9093) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CD3E at approximately 23 kDa. The expected band size for CD3E is at 23 kDa.

Figure 2. IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody (PB9093).
CD3 Epsilon was detected in paraffin-embedded section of human tonsil 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-CD3 Epsilon Antibody (PB9093) 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 CD3 Epsilon using anti-CD3 Epsilon antibody (PB9093).
CD3 Epsilon was detected in paraffin-embedded section of mouse spleen 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-CD3 Epsilon Antibody (PB9093) 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 CD3 Epsilon using anti-CD3 Epsilon antibody (PB9093).
CD3 Epsilon was detected in paraffin-embedded section of rat spleen 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-CD3 Epsilon Antibody (PB9093) 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 CD3 Epsilon using anti-CD3 Epsilon antibody (PB9093).
CD3 Epsilon was detected in frozen section of rat spleen tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD3 Epsilon Antibody (PB9093) 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. IHC analysis of CD3 Epsilon using anti-CD3 Epsilon antibody (PB9093).
CD3 Epsilon was detected in frozen section of mouse spleen tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD3 Epsilon Antibody (PB9093) 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 7. IF analysis of CD3E and CD68 using anti-CD3E antibody (PB9093) and anti-CD68 antibody (M00602)
CD3E and CD68 was detected in paraffin-embedded section of human tonsil 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-CD3E Antibody (PB9093) and mouse anti CD68 Antibody(M00602) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) and Biotin conjugated goat anti-rabbit IgG (BA1003) were used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 8. IF analysis of CD3E and CD20 using anti-CD3E antibody (PB9093) and anti-CD20 antibody (M03780-5).
CD3E and CD20 was detected in a paraffin-embedded section of human tonsil 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-CD3E antibody (PB9093) and mouse anti-CD20 antibody (M03780-5) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135), DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 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.