antibodies specific for human cd14 conjugated to alexa fluor 647, mouse mab, clone #m5e2,  (Novus Biologicals)

Journal: STAR Protocols

Article Title: Protocol for extracting and isolating porcine bone-marrow-derived macrophages from ribs

doi: 10.1016/j.xpro.2024.103085

Figure Lengend Snippet: Cell distribution and immunophenotypes of floating cells after 1-h selection (A) Optical images of cell culture before and after a 1-h selection. The scale bar is 50 μm. (B) Representative FACS plots of CD3, CD14, and CD45 in the cells before seeding (labeled in blue) and floating cells after 1 h of culture (labeled in red) ( N = 3 biological replicates). The graphs report the mean fluorescence intensity (MFI) of the three markers. An unpaired t-test was used as a statistical comparison method using GraphPad software. ∗ indicates p < 0.05, and ns indicates no significance ( p ≥ 0.05).

Article Snippet: CD14 Alexa Fluor 647 conjugated , Bio-Rad , MCA1218AF647.

Techniques: Selection, Cell Culture, Labeling, Fluorescence, Comparison, Software

Journal: STAR Protocols

Article Title: Protocol for extracting and isolating porcine bone-marrow-derived macrophages from ribs

doi: 10.1016/j.xpro.2024.103085

Figure Lengend Snippet: Cell morphology and immunophenotypes of BMDM after 24 h of stimulation (A) Representative optical image of BMDM after four days of culture. The scale bar is 50 μm. (B‒D) FACS plots and MFI analyses of CD14 (B), CD8086 (C), and SWC9 (D) in the groups of Control, M (LPS+IFNγ), M (IL-4+IL-13). The gray plot (B-D, in upper FACS plots) is the unstained group and is used for FACS gating. One-way ANOVA was used for statistical analysis. ∗Indicates p < 0.05, and ns indicates no significance ( p ≥ 0.05).

Article Snippet: CD14 Alexa Fluor 647 conjugated , Bio-Rad , MCA1218AF647.

Techniques:

Journal: STAR Protocols

Article Title: Protocol for extracting and isolating porcine bone-marrow-derived macrophages from ribs

doi: 10.1016/j.xpro.2024.103085

Figure Lengend Snippet:

Article Snippet: CD14 Alexa Fluor 647 conjugated , Bio-Rad , MCA1218AF647.

Techniques: Recombinant, Saline, Lysis, Isolation, SYBR Green Assay, Software, Sterility, Spectrophotometry, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Response to Oxidative Burst-Induced Hypoxia Is Associated With Macrophage Inflammatory Profiles as Revealed by Cellular Genome-Wide Association

doi: 10.3389/fimmu.2021.688503

Figure Lengend Snippet: Phenotypic characteristics of harvested cells after six days of in vitro incubation in serum-free media supplemented with recombinant bovine granulocyte-macrophage colony-stimulating factor. (A) Autofluorescence of Monocyte-Derived Macrophages (MDMs) (dark grey) versus monocyte (light grey), exited by blue laser and the emission was measured by 533/30 filter. (B) Expression of CD205 on MDMs. The harvested cells were stained with phytoerythrin (PE) conjugated anti-bovine CD-205 (dark grey) and compared against the unstained sample (light grey). (C) Expression of CD14 on MDMs. The harvested cells were stained with Alexa Flour 647 conjugated anti-human CD-14 (Clone TÜK4) (dark grey) versus the unstained sample (light grey). The cells were analyzed in BD Accuri™ C6 cytometer. The data from the flow cytometer were analyzed and graphed using FlowJo (v. 10).

Article Snippet: Harvested cells were labeled with RPE conjugated mouse anti-bovine CD205 (Clone: IL-A114, Bio-Rad, Mississauga, Ontario) and ALEXA FLUOR ® 647 conjugated mouse anti-human CD14 (Clone: TÜK4, Bio-Rad, Mississauga, Ontario), separately.

Techniques: In Vitro, Incubation, Recombinant, Derivative Assay, Expressing, Staining, Cytometry, Flow Cytometry