Anti-CD31/Pecam1 Antibody Picoband®

Western blot analysis of Pecam1 using anti-Pecam1 antibody (A01513-3).
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: rat liver tissue lysates,
Lane 2: rat lung tissue lysates,
Lane 3: mouse lung tissue 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-Pecam1 antigen affinity purified polyclonal antibody (Catalog # A01513-3) at 0.25 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 Pecam1 at approximately 120-130KD. The expected band size for Pecam1 is at 82KD.

IHC analysis of Pecam1 using anti-Pecam1 antibody (A01513-3).
Pecam1 was detected in paraffin-embedded section of human placenta 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-Pecam1 Antibody (A01513-3) 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 Pecam1 using anti-Pecam1 antibody (A01513-3).
Pecam1 was detected in paraffin-embedded section of mouse lung 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-Pecam1 Antibody (A01513-3) 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 Pecam1 using anti-Pecam1 antibody (A01513-3).
Pecam1 was detected in paraffin-embedded section of rat liver 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-Pecam1 Antibody (A01513-3) 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 Pecam1 using anti-Pecam1 antibody (A01513-3).
Pecam1 was detected in a paraffin-embedded section of human normal liver and alcoholic liver 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-Pecam1 Antibody (A01513-3) overnight at 4°C. DyLight 594 Donkey Anti-Mouse 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 fluorescence microscope and filter sets appropriate for the label used.

Construction of animal models and analysis of pathological damage. (A) TTC staining plots of the Sham and VD group. (B) Motion trajectory diagram. (C) Representative images stained with HE (scale bars = 50 μm). (D) Representative histopathological images obtained from TUNEL staining (scale bars = 50 μm), showing changes in the cortex hippocampus CA1. (E) Quantification of TUNEL+ cells in the hippocampal CA1 (n = 3). (F) Expression profile of CD31 between Sham and VD groups (qPCR). **P < 0.01. (G) Expression profile of HIF-1α between Sham and VD groups (qPCR; n = 6). ***P < 0.001. (H) WB strips. (I) Expression profile of CD31 between Sham and VD groups (WB). *P < 0.05. (J) Expression profile of HIF-1α between Sham and VD groups (WB; n = 3). ***P < 0.001.
Index in PubMed under a CC BY license. PMID: 40988927

Experimental validation of the key genes in vivo . (A) Representative images of immunofluorescence. (B–F) Expression profile of CCL2, VEGFA, SPP1, ANGPT2, and ANGPTL4 between Sham and VD groups (qPCR; n = 6). *P < 0.05; **P < 0.01; ***P < 0.001. (G) Immunofluorescence quantification of CD31. ***P < 0.001. (H) WB representative strips of the 5 key genes. (I–M) Protein expression level of CCL2, VEGFA, SPP1, ANGPT2, and ANGPTL4 between Sham and VD groups (WB; n = 3). *P < 0.05; **P < 0.01; ***P < 0.001.
Index in PubMed under a CC BY license. PMID: 40988927

Light flicker stimulation at 40 Hz does not increase angiogenesis in mice with rUCCAO. ( a , b ) CD31 immunohistochemical staining in the frontal lobe and hippocampus. Scale bar = 50 μm; the blue arrow indicates the positive area. ( c – f ) Positive area (%) and IOD of CD31 protein in the frontal lobe and hippocampus, respectively (n = 3/group. Means ± SEMs, *p < 0.05, n.s. not significant).
Index in PubMed under a CC BY license. PMID: 38049571