Anti-CD23/FCER2 Antibody Picoband™

Figure 1. Western blot analysis of CD23 using anti-CD23 antibody (PB9051).
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: rat thymus tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat PC-12 whole cell lysates,
Lane 4: mouse spleen tissue lysates,
Lane 5: mouse thymus tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse RAW264.7 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-CD23 antigen affinity purified polyclonal antibody (Catalog # PB9051) 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 CD23 at approximately 49 kDa. The expected band size for CD23 is at 36 kDa.

Figure 2. IHC analysis of CD23 using anti-CD23 antibody (PB9051).
CD23 was detected in paraffin-embedded section of Mouse Spleen 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-CD23 Antibody (PB9051) 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 CD23 using anti-CD23 antibody (PB9051).
CD23 was detected in paraffin-embedded section of Rat Spleen 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-CD23 Antibody (PB9051) 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 CD23 using anti-CD23 antibody (PB9051).
CD23 was detected in paraffin-embedded section of Human Tonsil 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-CD23 Antibody (PB9051) 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 CD23 using anti-CD23 antibody (PB9051).
CD23 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-CD23 Antibody (PB9051) 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. IF analysis of CD23 using anti-CD23 antibody (PB9051)
CD23 was detected in paraffin-embedded section of mouse lymphaden 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-CD23 Antibody (PB9051) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Figure 7. IF analysis of CD23 using anti-CD23 antibody (PB9051)
CD23 was detected in paraffin-embedded section of rat lymphaden 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-CD23 Antibody (PB9051) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Figure 8. Flow Cytometry analysis of mouse PBMC cells using anti-CD23 antibody (PB9051). Overlay histogram showing mouse PBMC cells stained with PB9051 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD23 Antibody PB9051,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 9. Flow Cytometry analysis of mouse BMC cells using anti-CD23 antibody (PB9051).
Overlay histogram showing mouse BMC cells stained with PB9051 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD23 Antibody (PB9051,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 10. Sandwich ELISA - Recombinant mouse CD23/FCER2 protein standard curve.
Use in combination with reagents from Mouse CD23/FCER2 ELISA Kit EZ-Set (DIY Antibody Pairs) (EZ0924).