mouse antihuman cd14 fitc monoclonal antibody  (Bio-Rad)

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction.

doi: 10.1152/ajpheart.00187.2016

Figure Lengend Snippet: Fig. 1. Flow cytometry gating of circulating leukocytes to isolate CD14 monocytes (MCs) and MC subsets based on CD62L expression. A: forward and side scatter area (FSC-A and SSC-A) plot of leukocytes. B: gating of single cells based on FSC width and FSC height. C: gating of live cells based on 7-amino-actinomycin D (7AAD) expression on the singlet population. D: gating for CD14 MCs from the live cell population. E: MC subsets were gated from the CD14 MCs into 2 populations based on CD62L and CD62L expression.

Article Snippet: After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.

Techniques: Flow Cytometry, Expressing

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction.

doi: 10.1152/ajpheart.00187.2016

Figure Lengend Snippet: Fig. 4. Mobilization of circulating leuko- cytes in peripheral blood of rabbits with chronic myocardial ischemia. Concentration ratio [(cells/ml on day of termination)/ (cells/ml at day 0)] of peripheral blood mononuclear cells (PBMCs; A), CD14 monocytes (B), CD14CD62L monocytes (C), and CD14CD62L (D) monocytes in peripheral blood of sham animals and rabbits with myocardial ischemia at different days after initiating chronic occlusion (CO). MCs, monocytes (sham n 3, 9-day CO n 4, 16-day CO n 4, 28-day CO n 5). *P 0.05.

Article Snippet: After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.

Techniques: Concentration Assay

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction.

doi: 10.1152/ajpheart.00187.2016

Figure Lengend Snippet: Fig. 5. Infiltration of autologous monocytes into ischemic myocardium. A: confocal microscopy image of autologous monocytes (all CD14 monocytes, blue) infiltrated in midmyocardial region of ischemic myocardium (16 days CO), with CD31 labeled (red) endothelium. Arrows indicate large vessel running parallel to the image, whereby monocyte are clustered around the vessel. B: temporal changes in relative tissue volume containing (%) CD14CD62L and CD14CD62L monocytes in myocardium of sham animals and rabbits with myocardial ischemia. C: relative vascular volume density (%) was determined as the number of voxels corresponding to vascular cast that were located within the monocyte occupied region (CD14CD62L or CD14CD62L) in ischemic myocardium. CO, initiation of chronic occlusion; MC, monocyte (sham n 3, 9-day CO n 4, 16-day CO n 4, 28-day CO n 5). *P 0.05.

Article Snippet: After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.

Techniques: Confocal Microscopy, Labeling

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction.

doi: 10.1152/ajpheart.00187.2016

Figure Lengend Snippet: Fig. 7. Quantification of the transmural dis- tribution of monocyte subsets in ischemic myocardium. Transmural distribution of monocyte subsets (CD14CD62L vs. CD14 CD62L) sham (A), 9-day CO (B), 16-day CO (C), or 28-day CO (D). CO: initiation of chronic occlusion; ENDO, endocardium; MC, monocyte; MID, midmyocardium; EPI, epicardium (sham n 3, 9-day CO n 4, 16-day CO n 4, 28-day CO n 5). *P 0.05, **P 0.01, ***P 0.001.

Article Snippet: After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.

Techniques:

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction.

doi: 10.1152/ajpheart.00187.2016

Figure Lengend Snippet: Fig. 8. Phenotypic and functional compari- son of rabbit monocyte subsets based on adhesion characteristics. A: phase contrast microscopy images of adherent CD14 CD62L (left) or CD14CD62L (right) monocytes in the presence of tissue culture plastic or fibronectin (FN) coated tissue cul- ture plastic. B: quantification of the number of adherent CD14CD62L or CD14 CD62L monocytes in the presence of tissue culture plastic or fibronectin (FN)-coated tis- sue culture plastic. CD14CD62L mono- cytes showed significantly greater adhesive capacity. C: quantification of the cell surface area of adherent CD14CD62L or CD14 CD62L monocytes in the presence of tissue culture plastic or FN-coated tissue culture plastic. CD14CD62L monocytes were significantly larger than CD14CD62L monocytes in the presence of tissue culture plastic. *P 0.05, **P 0.01; n 3 rabbits, conducted in duplicate.

Article Snippet: After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.

Techniques: Functional Assay, Microscopy, Adhesive

Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction.

doi: 10.1152/ajpheart.00187.2016

Figure Lengend Snippet: Fig. 9. Cell surface receptor expression and migration properties of monocyte subsets. Percentage of CD14CD62L and CD14 CD62L monocytes expressing CCR2 (A) and CXCR4 (B) (n 4). Significantly greater percentage of CD14CD62L monocytes express the chemokine receptor CCR2. C: Transwell chemotaxis migration assay exam- ining relative migration of monocyte subsets towards various chemoattractants (MCP1, fMLP, SDF1). Significantly greater number of CD14CD62L monocytes migrate to- wards MCP1 (n 5). CCR2, C-C chemokine receptor 2; CXCR4, C-X-C chemokine re- ceptor type 4; fMLP, 10-8M N-formyl-Met- Leu-Phe; MCP1, monocyte chemoattractant protein 1; SDF1, stromal derived factor 1. *P 0.05.

Article Snippet: After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.

Techniques: Cell Surface Receptor Assay, Expressing, Migration, Chemotaxis Assay, Derivative Assay