Anti-PAR1/Thrombin Receptor/F2R Antibody Picoband®

Western blot analysis of F2R using anti-F2R antibody (A03352-1).
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: human placenta tissue lysates,
Lane 2: human U-87MG whole cell lysates.
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-F2R antigen affinity purified polyclonal antibody (Catalog # A03352-1) 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 F2R at approximately 47KD. The expected band size for F2R is at 47KD.

(A) The binding energy between the six most important targets and the core compounds was calculated via molecular docking models. (B) Piperlongumine and PTGS2, binding energy = −8.56 kcal/mol. (C) Piperlongumine and F2R, binding energy = −8.81 kcal/mol. (D) Piperine and F2R, binding energy = −8.74 kcal/mol.
Index in PubMed under a CC BY license. PMID: 38328576

Cytokine levels in the knee cartilage tissue of 100 and 300 mg/kg PLE-treated rats. (A–L) mRNA expression of F2R, F3, IL-1β, IL-6, IL-17A, MMP-1, MMP-2, MMP-3, MMP-9, MMP-13, NOS2, PTGS2, PGE2, and TNF-α determined by qRT-PCR. (M) WB analysis of the protein expression of F2R, F3, IL-17A, MMP-1, MMP-2, MMP-9, and PTGS2. ### p < 0.001 vs. NT, ** p < 0.05 vs. MIA, *** p < 0.001 vs. MIA. NT: non-treated, INDO 3: indomethacin 3 mg/kg.
Index in PubMed under a CC BY license. PMID: 38328576

IHC analysis of F2R using anti-F2R antibody (A03352-1).
F2R was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-F2R Antibody (A03352-1) 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 F2R using anti-F2R antibody (A03352-1).
F2R was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-F2R Antibody (A03352-1) 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.

Flow Cytometry analysis of HL-60 cells using anti-F2R antibody (A03352-1).
Overlay histogram showing HL-60 cells stained with A03352-1 (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-F2R Antibody (A03352-1, 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.