Anti-Cd86 Antibody Picoband®

YNJ reduces the secretion of inflammatory factors and promotes M2 macrophage polarization. (A–C) ELISA of TNF-α (A) , IL-6 (B) , and IL-1β (C) contents in the supernatant of cultured RAW264.7 macrophages. (D) IF images of CD86 and CD206 detection in cultured RAW264.7 macrophages. (E, F) Statistical analysis of CD86 and CD206 expression by IF. The data are presented as the mean ± SD (n = 6). ** P < 0.01.
Index in PubMed under a CC BY license. PMID: 39925847

Western blot analysis of Cd86 using anti-Cd86 antibody (A00220-4).
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 lung tissue lysates,
Lane 2: mouse lung tissue 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-Cd86 antigen affinity purified polyclonal antibody (Catalog # A00220-4) 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 Cd86 at approximately 60-80 kDa. The expected band size for Cd86 is at 35 kDa.

IHC analysis of Cd86 using anti-Cd86 antibody (A00220-4).
Cd86 was detected in a paraffin-embedded section of rat spleen 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-Cd86 Antibody (A00220-4) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of Cd86 using anti-Cd86 antibody (A00220-4). Cd86 was detected in a paraffin-embedded section of human normal and preterm placenta 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 1;200 rabbit anti-Cd86 Antibody (A00220-4) overnight at 4°C. DyLight 594 Goat Anti-Rabbit IgG (H+L) was used as secondary antibody at 1:500 dilution and incubated for 45 minutes at 37°C. The section was counterstained with DAPI. Visualize using a Leica DM2500 system.

Flow Cytometry analysis of ANA-1 cells using anti-Cd86 antibody (A00220-4).
Overlay histogram showing ANA-1 cells stained with A00220-4 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Cd86 Antibody (A00220-4, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.