Anti-CD23/FCER2 Antibody Picoband®

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.

FB cells in pSS model mice. (A) F (CD23 + CD21 − ) and MZ (CD23 − CD21 + ) B cells among the CD19 + cells identified in the spleen, the salivary glands, and the lungs of 10-week-old control and pSS model. Representative results are shown. (B) Proportions and numbers of follicular B cells in the spleen of 10-week-old control mice and pSS model mice. Data are presented as mean ± SD of four to five mice per group. (C) Proportions and numbers of follicular B cells in the salivary gland tissues of 12-week-old control and SS model mice. Data are presented as mean ± SD of four mice per group. (D) Population of follicular B cells in the lungs of 8- and 16-week-old control mice and SS model mice. Data are presented as mean ± SD of three to eight mice per group. * p < 0.05, ** p < 0.01. (E) CD23 + cells were detected through immunohistochemical analysis by using lung sections of 8-week-old control and SS model mice. Scale bar: 100 μm. (F) B220 + B cells and CD23 + cells were detected through immunofluorescence analysis by using lung sections obtained from 8-week-old control and pSS model mice. Nuclei were stained with DAPI. Scale bar: 50 μm.
Index in PubMed under a CC BY license. PMID: 37475871

CD23 + B-cell differentiation via IL-4 in the lungs of the pSS model mice. (A) Il4 mRNA expressions were analyzed through qRT-PCR, by using spleen and lung tissues of 8-week-old control and SS model mice. Data are presented as mean ± SD of four mice per group. * p < 0.05. (B) Gata3 (upper panel) and Tbx21 (lower panel) mRNA expressions were analyzed through qRT-PCR, by using spleen, salivary gland, and lung tissues of 8-week-old control and SS model mice. Data are presented as mean ± SD of three to four mice per group. * p < 0.05. (C) CD19 + B cells isolated from the lungs of control and SS model mice were stimulated in vitro with an anti-CD40 mAb (5 µg/ml) and recombinant IL-4 (100 ng/ml) for 7 days. The relative cell number of CD23 + B cells to the unstimulated cells was evaluated. Data are presented as mean ± SD of triplicates per group. * p < 0.05, ** p <0.01.
Index in PubMed under a CC BY license. PMID: 37475871

CD23 + FB cell differentiation within the lungs of anti-CD4 mAb-treated pSS model mice. (A) Anti-CD4 mAb was intraperitoneally administered to pSS model mice between their sixth to eighth week of their lives. We assessed the proportions of CD4 + T cells in the spleen and of CD4 + T and CD19 + B cells in the lungs of isotype control mAb-treated and of anti-CD4 mAb-treated pSS model mice. Data are presented as mean ± SD of seven mice per group. * p < 0.05. (B) Number of CD4 + T cells in the spleen and of CD4 + T and CD19 + B cells in the lungs of isotype control mAb- and of anti-CD4 mAb-treated pSS model mice. Data are presented as mean ± SD of seven mice per group. ** p < 0.01. (C) Pulmonary lesions in anti-CD4 mAb-treated pSS model mice were histologically evaluated. Representative images of HE-stained lung tissues sections of isotype control mAb-treated and anti-CD4 mAb-treated pSS model mice. Scale bar: 100 μm. (D) The number of foci in the pulmonary lesions was counted by using HE-stained sections. Data are presented as mean ± SD of seven mice per group. (E) CD23 + cells in the pulmonary lesions were evaluated immunohistochemically. Representative images are shown for each group. Scale bar: 100 μm. (F) CD23 + CD19 + FB cells and CD23 − CD19 + B cells were evaluated through flow cytometric analysis by using lung tissues. Representative results are shown for each group. (G) The proportions of CD23 + CD19 + FB cells and of CD23 − CD19 + B cells were analyzed through flow cytometry. Data are presented as mean ± SD of seven mice per group. * p < 0.05.
Index in PubMed under a CC BY license. PMID: 37475871

Preventive effect of the anti-CD4 mAb in the pulmonary lesions of pSS model mice. (A) Anti-CD4 mAb was intraperitoneally administered to pSS model mice between their fourth to sixth week of their lives. Pulmonary lesions in anti-CD4 mAb-treated pSS model mice were histologically evaluated. Representative images of HE-stained lung tissues sections of isotype control mAb-treated and anti-CD4 mAb-treated pSS model mice. Scale bar: 100 μm. (B) The area of foci in the pulmonary lesions was measured by using HE-stained sections. Data are presented as mean ± SD of seven to eight mice per group. ** p <0.01. (C) We assessed the proportions of CD4 + T cells in the spleen and lung of isotype control mAb-treated and of anti-CD4 mAb-treated pSS model mice. Data are presented as mean ± SD of seven to eight mice per group. ** p < 0.01. (D) Number of CD4 + T, CD19 + B, and CD19 + CD23 + FB cells in the lung of isotype control mAb-treated and of anti-CD4 mAb-treated pSS model mice. Data are presented as mean ± SD of seven to eight mice per group. ** p <0.01.
Index in PubMed under a CC BY license. PMID: 37475871

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.

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.

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.

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.

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.

Flow Cytometry analysis of mouse PBMC cells using anti-CD23 antibody (PB9051).
Overlay histogram showing mouse PBMC cells stained with PB9051 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

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.

Flow Cytometry analysis of mouse BMC cells using anti-CD23 antibody (PB9051).
Overlay histogram showing mouse BMC cells stained with PB9051 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

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).