mouse anti cd86 primary antibody  (Abcam)

Journal: Bioactive Materials

Article Title: A self-locking conductive cardiac patch for immediate electrical integration with infarcted rat myocardium

doi: 10.1016/j.bioactmat.2025.10.045

Figure Lengend Snippet: The effects of patches on angiogenesis and inflammation in infarcted tissues 4 weeks post-MI. a) Immunofluorescence staining of α-smooth muscle actin (α-SMA, a vascular protein marker, green) and von Willebrand factor (vWF, an endothelial marker, red) of the infarct region in the MI, BMN-P, BMN-CP and CBMN-CP groups. b, c) Quantitative analysis of α-SMA (b) and vWF (c) in different groups based on fluorescent staining images (n = 4). d) Representative immunofluorescence staining of the infarct region in different groups for CD86 (green) and CD206 (red). e, f) Fluorescence intensity statistics of CD86 (e) and CD206 (f) in different groups based on fluorescent staining images (n = 4). Nuclei are stained blue with DAPI. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: The tissue sections were then incubated with the following primary antibodies: rabbit Connexin 43 primary antibody (BOSTER, BA1727, 1:200, China), mouse α-actinin primary antibody (Abcam, AB9465, 1:200, UK), mouse α-SMA primary antibody (Wuhan Sanying, 67735-1-IG, 1:400, China), rabbit vWF primary antibody (Wuhan Sanying, 27186-1-AP, 1:300, China), mouse CD86 primary antibody (BOSTER, BA4121, 1:100, China) and rabbit CD206 primary antibody (Wuhan Sanying, 18704-1-AP, 1:400, China).

Techniques: Immunofluorescence, Staining, Marker, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Vascular endothelial growth factor attenuates neointimal hyperplasia of decellularized small-diameter vascular grafts by modulating the local inflammatory response

doi: 10.3389/fbioe.2022.1066266

Figure Lengend Snippet: Representative immunofluorescence images of subcutaneous implants at day 28 in each group (A) . CD68 and CD206 stained in green, CD86 stained in red. Quantification of CD68 + total macrophages, CD86 + M1-macrophages, CD206+ M2-macrophages and M2/M1 (CD206+/CD86+) ratio (B) . * p < 0.05. ** p < 0.01. ns represents no significant difference.

Article Snippet: M1 macrophages were stained with mouse anti-CD86 primary antibody (1:100, ab220188, Abcam, United States).

Techniques: Immunofluorescence, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Vascular endothelial growth factor attenuates neointimal hyperplasia of decellularized small-diameter vascular grafts by modulating the local inflammatory response

doi: 10.3389/fbioe.2022.1066266

Figure Lengend Snippet: Representative immunofluorescence images of vascular grafts at day 28 in each group (A) . CD68 and CD206 stained in green, CD86 stained in red. Quantification of CD68 + total macrophages, CD86 + M1-macrophages, CD206+ M2-macrophages and M2/M1 (CD206+/CD86 + ) ratio (B) . * p < 0.05. ** p < 0.01. ns represents no significant difference.

Article Snippet: M1 macrophages were stained with mouse anti-CD86 primary antibody (1:100, ab220188, Abcam, United States).

Techniques: Immunofluorescence, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Vascular endothelial growth factor attenuates neointimal hyperplasia of decellularized small-diameter vascular grafts by modulating the local inflammatory response

doi: 10.3389/fbioe.2022.1066266

Figure Lengend Snippet: Representative immunofluorescence images of vascular grafts at day 7 in each group (A) . CD68 and CD206 stained in green, CD86 stained in red. Quantification of CD68 + total macrophages, CD86 + M1-macrophages, CD206+ M2-macrophages and M2/M1 (CD206+/CD86 + ) ratio (B) . * p < 0.05. ** p < 0.01. ns represents no significant difference.

Article Snippet: M1 macrophages were stained with mouse anti-CD86 primary antibody (1:100, ab220188, Abcam, United States).

Techniques: Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: A novel peptide that improves metabolic parameters without adverse central nervous system effects

doi: 10.1038/s41598-017-13690-9

Figure Lengend Snippet: Quantitative analyses in inguinal adipose tissue of DIO rats. Panels a–d: Immunohistochemical representative images (400x magnification, scale bar = 50 µm) showing specific UCP1-immunostained cells from the inguinal white adipose tissue of DIO Wistar rats (oral treatments: panel a, saline; panel b, Pep19, 100 μg/Kg; panel c, Pep19, 300 μg/Kg; panel d, Pep19, 600 μg/Kg). Quantitative analyses of UCP1 labeled cells were determined from at least 25 different fields in each slice (n = 8–10), and examined using ImageJ software at 400x magnification; results are expressed as number of UCP1 positive cells per mm 2 . Representative UCP1-immunostained cells (fast red dye) obtained from each treatment are indicated by arrows. Panel e: Quantitative analyses suggest that oral administration of Pep19 600 μg/Kg increases the number of UCP1-immunostained cells in the inguinal adipose tissue compared to saline or other treatments (n = 8–10; *p < 0.05, saline vs Pep19 600 μg/Kg). Note that in Pep19 treated animals (panels, c and d) immunostained-UCP1 cells seem to occur in fibrotic-like areas of the adipose tissue. Panel f: Quantitative Western blot analysis suggested similar levels of UCP1 expression in either saline or Pep19 (600 μg/Kg) treated groups of DIO animals; (each line shown on the upper panel f, is representative of one individual animal treated with either saline or Pep19 600 μg/Kg; n = 7). Note that the ratio of UCP1 negative cells to positive cells is fairly large and this could contribute to lack in change in the protein levels seen in the Western Blots (panel f). Panel g: Quantitative analyses of the number of adipocytes (adipocytes/µm 2 ; 500–800 cells were counted per animal (n = 3), in 14–18 different fields of the H&E stained slices of adipose tissue from animals treated with either saline or Pep19 (600 μg/Kg; *p < 0.05). Panel h: Quantitative analyses of the adipocyte area were performed measuring 15 different adipocytes in each of the 6 fields analyzed per H&E stained slices from each animal (n = 3), using H&E stained slices of adipose tissue from animals treated with either saline or Pep19 (600 μg/Kg; *p < 0.01). All results are expressed as the means ± standard error of the mean (SEM). The statistical comparisons were performed using Student’s t-test or analysis of variance (ANOVA), followed by ad-hoc Tukey’s test using GraphPad Prism software. Crude Western blot membranes are shown on Supplemental material.

Article Snippet: Membranes were blocked for 1 h at room temperature in Tris-buffered saline Tween-20 buffer with 5% nonfat dry milk, followed by overnight incubation at 4 °C with specific mouse primary antibodies for UCP1 (Abcam, ab23841) or GAPDH (sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in Tris-buffered saline with 5% nonfat dry milk.

Techniques: Immunohistochemical staining, Labeling, Software, Western Blot, Expressing, Staining

Journal: Scientific Reports

Article Title: A novel peptide that improves metabolic parameters without adverse central nervous system effects

doi: 10.1038/s41598-017-13690-9

Figure Lengend Snippet: Signaling pathways induced by Pep19 in 3T3-L1 adipocyte cells. Panels a and b: Relative UCP1 expression in 3T3-L1 adipocyte cells. Panel a: Cells were exposed to rosiglitazone (RSG, 5µM), or different concentrations of hemopressin (HP, 0.1–10 µM) or Pep19 (0.1–10 µM). Panel b: 3T3-L1 adipocyte cells exposed to Pep19 (1 µM) for 24 h in the absence or presence of either RSG (5 µM), the CB1R agonist WIN55,212–2 (1 µM), the CB1R antagonist AM251 (1 µM) or the CB1R inverse agonist HP (1 µM). Western blots were conducted using mouse anti-UCP1 antibodies, and anti-GAPDH antibodies were used as loading controls. Panels c and d: 3T3-L1 cells were starved for 16 h in serum-free medium prior to stimulation (vehicle or Pep19, 1 μM) for the indicated time period. Western blots were carried out using: Panel c, mouse monoclonal anti-phosphoERK1/2, and rabbit polyclonal anti-total ERK1/2; Panel d, rabbit phospho-AKT S473 (anti-pAKT, S473) and mouse monoclonal anti-tubulin antibodies. Imaging and band intensity measurements were performed using the Odyssey imaging system (LI-COR, Lincoln, NE) according to the manufacturer’s protocols. Data are representative of three independent experiments that produced similar results. Unt, cells not treated with vehicle (Veh) or peptide 19 (Pep19). The statistical comparisons were performed using Student’s t-test or analysis of variance (ANOVA), followed by ad-hoc Tukey’s test using GraphPad Prism software * p < 0.05; ** p < 0.001. Crude Western blot membranes are shown on Supplemental material.

Article Snippet: Membranes were blocked for 1 h at room temperature in Tris-buffered saline Tween-20 buffer with 5% nonfat dry milk, followed by overnight incubation at 4 °C with specific mouse primary antibodies for UCP1 (Abcam, ab23841) or GAPDH (sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in Tris-buffered saline with 5% nonfat dry milk.

Techniques: Expressing, Western Blot, Imaging, Produced, Software

Journal: Scientific Reports

Article Title: A novel peptide that improves metabolic parameters without adverse central nervous system effects

doi: 10.1038/s41598-017-13690-9

Figure Lengend Snippet: Quantitative analyses in inguinal adipose tissue of DIO rats. Panels a–d: Immunohistochemical representative images (400x magnification, scale bar = 50 µm) showing specific UCP1-immunostained cells from the inguinal white adipose tissue of DIO Wistar rats (oral treatments: panel a, saline; panel b, Pep19, 100 μg/Kg; panel c, Pep19, 300 μg/Kg; panel d, Pep19, 600 μg/Kg). Quantitative analyses of UCP1 labeled cells were determined from at least 25 different fields in each slice (n = 8–10), and examined using ImageJ software at 400x magnification; results are expressed as number of UCP1 positive cells per mm 2 . Representative UCP1-immunostained cells (fast red dye) obtained from each treatment are indicated by arrows. Panel e: Quantitative analyses suggest that oral administration of Pep19 600 μg/Kg increases the number of UCP1-immunostained cells in the inguinal adipose tissue compared to saline or other treatments (n = 8–10; *p < 0.05, saline vs Pep19 600 μg/Kg). Note that in Pep19 treated animals (panels, c and d) immunostained-UCP1 cells seem to occur in fibrotic-like areas of the adipose tissue. Panel f: Quantitative Western blot analysis suggested similar levels of UCP1 expression in either saline or Pep19 (600 μg/Kg) treated groups of DIO animals; (each line shown on the upper panel f, is representative of one individual animal treated with either saline or Pep19 600 μg/Kg; n = 7). Note that the ratio of UCP1 negative cells to positive cells is fairly large and this could contribute to lack in change in the protein levels seen in the Western Blots (panel f). Panel g: Quantitative analyses of the number of adipocytes (adipocytes/µm 2 ; 500–800 cells were counted per animal (n = 3), in 14–18 different fields of the H&E stained slices of adipose tissue from animals treated with either saline or Pep19 (600 μg/Kg; *p < 0.05). Panel h: Quantitative analyses of the adipocyte area were performed measuring 15 different adipocytes in each of the 6 fields analyzed per H&E stained slices from each animal (n = 3), using H&E stained slices of adipose tissue from animals treated with either saline or Pep19 (600 μg/Kg; *p < 0.01). All results are expressed as the means ± standard error of the mean (SEM). The statistical comparisons were performed using Student’s t-test or analysis of variance (ANOVA), followed by ad-hoc Tukey’s test using GraphPad Prism software. Crude Western blot membranes are shown on Supplemental material.

Article Snippet: Next, membranes were incubated overnight at 4 °C with specific mouse primary antibodies anti-UCP1 (Abcam, ab23841) or GAPDH (sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in Tris-HCl-buffered saline, pH 7.4, containning 5% nonfat dry milk.

Techniques: Immunohistochemical staining, Labeling, Software, Western Blot, Expressing, Staining

Journal: Scientific Reports

Article Title: A novel peptide that improves metabolic parameters without adverse central nervous system effects

doi: 10.1038/s41598-017-13690-9

Figure Lengend Snippet: Signaling pathways induced by Pep19 in 3T3-L1 adipocyte cells. Panels a and b: Relative UCP1 expression in 3T3-L1 adipocyte cells. Panel a: Cells were exposed to rosiglitazone (RSG, 5µM), or different concentrations of hemopressin (HP, 0.1–10 µM) or Pep19 (0.1–10 µM). Panel b: 3T3-L1 adipocyte cells exposed to Pep19 (1 µM) for 24 h in the absence or presence of either RSG (5 µM), the CB1R agonist WIN55,212–2 (1 µM), the CB1R antagonist AM251 (1 µM) or the CB1R inverse agonist HP (1 µM). Western blots were conducted using mouse anti-UCP1 antibodies, and anti-GAPDH antibodies were used as loading controls. Panels c and d: 3T3-L1 cells were starved for 16 h in serum-free medium prior to stimulation (vehicle or Pep19, 1 μM) for the indicated time period. Western blots were carried out using: Panel c, mouse monoclonal anti-phosphoERK1/2, and rabbit polyclonal anti-total ERK1/2; Panel d, rabbit phospho-AKT S473 (anti-pAKT, S473) and mouse monoclonal anti-tubulin antibodies. Imaging and band intensity measurements were performed using the Odyssey imaging system (LI-COR, Lincoln, NE) according to the manufacturer’s protocols. Data are representative of three independent experiments that produced similar results. Unt, cells not treated with vehicle (Veh) or peptide 19 (Pep19). The statistical comparisons were performed using Student’s t-test or analysis of variance (ANOVA), followed by ad-hoc Tukey’s test using GraphPad Prism software * p < 0.05; ** p < 0.001. Crude Western blot membranes are shown on Supplemental material.

Article Snippet: Next, membranes were incubated overnight at 4 °C with specific mouse primary antibodies anti-UCP1 (Abcam, ab23841) or GAPDH (sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:1000 in Tris-HCl-buffered saline, pH 7.4, containning 5% nonfat dry milk.

Techniques: Expressing, Western Blot, Imaging, Produced, Software