mouse antihuman monoclonal cd14-fitc  (Bio-Rad)

Journal: Journal of cellular immunology

Article Title: Preliminary Evidence of Differentially Induced Immune Responses by Microparticle-adsorbed LPS in Patients with Crohn’s Disease

doi: 10.33696/immunology.4.152

Figure Lengend Snippet: Stimulation of PBMC with LPS, TiO 2 and CaCL 2 dose response curves in Crohn’s disease and healthy controls, as individual components of the LPS-TiO2-Ca 2+ conjugate.

Article Snippet: Following culture, cells were washed twice in TCM, prior to staining with mouse antihuman monoclonal CD14-FITC (Serotec, UK) in the residual cell suspension (200 μl cells in TCM) for 30 mins on ice in the dark.

Techniques:

Journal: Animals : an open access journal from MDPI

Article Title: MHCII Expression on Peripheral Blood Monocytes in Canine Lymphoma: An Impact of Glucocorticoids.

doi: 10.3390/ani12162135

Figure Lengend Snippet: Figure 1. Representative dot plots depicting the gating strategy of canine peripheral blood leukocytes. (A) Forward scatter (FSC-hight) vs. side scatter (SSC-hight) dot plot showing leukocyte populations (population colors result from the regions set on the cytogram C); (B) debris and non-cellular elements were eliminated by setting a region defining CD45+ leukocytes (R1) on the SSC vs. CD45:APC dot plot; (C) leukocyte populations “(R1)” were analyzed on the CD45:APC/CD14:FITC dot plot in order to separate CD14+ monocytes (R2—red color) from lymphocytes (R3—blue color).

Article Snippet: Samples were labeled with the following panel of monoclonal antibodies: rat anti-dog CD45 APC-conjugated (clone YKIX716.13, Bio-Rad, Hercules, CA, USA), mouse anti-dog CD11b unconjugated (clone CA16.3E10, Bio-Rad), rat anti-dog MHC class II FITC-conjugated (clone YKIX334.2; Bio-Rad) and mouse antihuman CD14 FITC or Alexa Fluor®647-conjugated (clone TÜK4; Bio-Rad) that cross-reacts with canine CD14 molecules [21].

Techniques:

Journal: Animals : an open access journal from MDPI

Article Title: MHCII Expression on Peripheral Blood Monocytes in Canine Lymphoma: An Impact of Glucocorticoids.

doi: 10.3390/ani12162135

Figure Lengend Snippet: Figure 2. Representative dot plots depicting gating strategy of peripheral blood monocytes of dog with lymphoma. (A) Regions were set for all CD11b+ myeloid cells (R1) and for CD14+ monocytes (R2) on the CD14:APC vs. CD11b:PE double fluorescence dot plot; (B) distribution of all leukocytes (CD11b+ cells—red color) and (C) monocytes (from the R2 region—blue color) according to the expression of CD14 and MHCII. Bottom-row dot plots show isotype controls, separate for each Ab: anti-CD14 (A,B) and anti-MHCII (C). Controls allowed us to set the quadrants.

Article Snippet: Samples were labeled with the following panel of monoclonal antibodies: rat anti-dog CD45 APC-conjugated (clone YKIX716.13, Bio-Rad, Hercules, CA, USA), mouse anti-dog CD11b unconjugated (clone CA16.3E10, Bio-Rad), rat anti-dog MHC class II FITC-conjugated (clone YKIX334.2; Bio-Rad) and mouse antihuman CD14 FITC or Alexa Fluor®647-conjugated (clone TÜK4; Bio-Rad) that cross-reacts with canine CD14 molecules [21].

Techniques: Expressing

Journal: Animals : an open access journal from MDPI

Article Title: MHCII Expression on Peripheral Blood Monocytes in Canine Lymphoma: An Impact of Glucocorticoids.

doi: 10.3390/ani12162135

Figure Lengend Snippet: Figure 3. (A) The percentage of CD11b+CD14+ monocytes: MHCII+ and MHCII−, in the peripheral blood of healthy dogs (n = 11) and dogs with lymphoma: not receiving any drugs (NRG, n = 10) and receiving glucocorticoids (RG, n = 8). Arithmetic mean ± standard deviation. Significant differences between the percentage of MHCII+ and MHCII−monocytes among each group: # p < 0.05, ### p ≤0.001 (Wilcoxon) and for the percentage of MHCII+ or MHCII−monocytes in comparison to healthy dogs: *** p ≤0.001 (no significant differences between dogs with lymphoma NRG and RG) (Kruskal–Wallis and post hoc Dunn analysis). (B) The results are shown as the ratio of CD11b+CD14+ monocyte percentages: MHCII+ to MHCII−. Arithmetic mean ± standard deviation. Significant differences between groups: *** p ≤0.001 (Kruskal–Wallis and post hoc Dunn analysis).

Article Snippet: Samples were labeled with the following panel of monoclonal antibodies: rat anti-dog CD45 APC-conjugated (clone YKIX716.13, Bio-Rad, Hercules, CA, USA), mouse anti-dog CD11b unconjugated (clone CA16.3E10, Bio-Rad), rat anti-dog MHC class II FITC-conjugated (clone YKIX334.2; Bio-Rad) and mouse antihuman CD14 FITC or Alexa Fluor®647-conjugated (clone TÜK4; Bio-Rad) that cross-reacts with canine CD14 molecules [21].

Techniques: Standard Deviation, Comparison

Journal: Animals : an open access journal from MDPI

Article Title: MHCII Expression on Peripheral Blood Monocytes in Canine Lymphoma: An Impact of Glucocorticoids.

doi: 10.3390/ani12162135

Figure Lengend Snippet: Figure 4. The number of CD14+ monocytes in the peripheral blood of healthy dogs (n = 11) and dogs with lymphoma: not receiving any drugs (NRG, n = 10) and receiving glucocorticoids (RG, n = 8). (A) Total number of CD14+ monocytes per microliter of peripheral blood. The number of CD14+ cells was calculated based on their percentage in relation to all leukocytes (CD45+ cells) and white blood cell count. (B) The number of MHCII+ and (C) MHCII−monocyte subsets. Arithmetic mean ± standard deviation. Significant differences between groups: * p < 0.05, ** p ≤0.01, *** p ≤0.001 (Kruskal–Wallis and Dunn post hoc analysis). (D) The numbers of all evaluated cell populations in individual patients: MHCII−and MHCII+ monocytes as well as all CD14+ cell population.

Article Snippet: Samples were labeled with the following panel of monoclonal antibodies: rat anti-dog CD45 APC-conjugated (clone YKIX716.13, Bio-Rad, Hercules, CA, USA), mouse anti-dog CD11b unconjugated (clone CA16.3E10, Bio-Rad), rat anti-dog MHC class II FITC-conjugated (clone YKIX334.2; Bio-Rad) and mouse antihuman CD14 FITC or Alexa Fluor®647-conjugated (clone TÜK4; Bio-Rad) that cross-reacts with canine CD14 molecules [21].

Techniques: Cell Counting, Standard Deviation

Journal: Frontiers in Immunology

Article Title: IL-36γ Is a Strong Inducer of IL-23 in Psoriatic Cells and Activates Angiogenesis

doi: 10.3389/fimmu.2018.00200

Figure Lengend Snippet: (A) IL-36 receptor (IL-36R) (red) on macrophages; CD14+ (green) (magnification 20×). (B) Following 48 h stimulation with IL-36γ, IL-1α, IL-17A, TNFα, and IFNγ (24 h priming), supernatant was analysed by ELISA for TNFα and IL-23. Unpaired t -test * p < 0.05, ** p < 0.01 psoriasis versus healthy (sample size: psoriasis = 9, healthy = 9, and boiled control n = 3).

Article Snippet: CD14+ purity was tested by FACs analysis with mouse antihuman CD14 FITC conjugated or mouse IgG isotype control (both 1:100; both ImmunoTools, Friesoythe, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Immunology

Article Title: IL-36γ Is a Strong Inducer of IL-23 in Psoriatic Cells and Activates Angiogenesis

doi: 10.3389/fimmu.2018.00200

Figure Lengend Snippet: Human umbilical vein endothelial cell monolayer was stimulated with or without TNFα 10 ng for 24 h. 1 × 10 5 monocytes were allowed to adhere to the monolayer for 30 min. Cells were visualised by immunofluorescence microscopy following CD14+ staining (B) and counted (A) (patient monocytes: psoriasis = 8; healthy = 8). Magnification 40×. Unpaired t -test * p < 0.05 and ** p < 0.01.

Article Snippet: CD14+ purity was tested by FACs analysis with mouse antihuman CD14 FITC conjugated or mouse IgG isotype control (both 1:100; both ImmunoTools, Friesoythe, Germany).

Techniques: Immunofluorescence, Microscopy, Staining

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