cd14 pe af750  (Bio-Rad)

Journal: bioRxiv

Article Title: Gene expression profiling of bovine raw milk is linked to the inflammatory status of the udder

doi: 10.1101/2025.09.03.671445

Figure Lengend Snippet: Cells from three milk samples were collected by centrifugation, labelled with antibodies against CD3 (T lymphocytes), CD14 (macrophages) and G1 (neutrophils) and sorted on a MoFlo Astrios EQ cell sorter. RNA was extracted from purified cell populations and analyzed by high-throughput RT-qPCR. Gene expression values were calculated against a theoretical control sample and analyzed by principal component analysis (A) and hierarchical clustering (B).

Article Snippet: Antibodies used in the present study were CD14-PE-AF750 (Bio-Rad, reference MCA1568P750, clone Tük4), CD45-PE (Bio-Rad, reference MCA2220PE, clone 1.11.32), G1 (Kingfisher Biotech, reference WSC0608B-100, clone CH138A) and CD3 (Kingfisher Biotech, reference WS0561B-100, clone MM1A).

Techniques: Centrifugation, Purification, High Throughput Screening Assay, Quantitative RT-PCR, Gene Expression, Control

Journal: bioRxiv

Article Title: Gene expression profiling of bovine raw milk is linked to the inflammatory status of the udder

doi: 10.1101/2025.09.03.671445

Figure Lengend Snippet: Milk samples were aseptically collected from 38 individual quarters. Milk was mixed with one volume of DPBS and cells were recovered by centrifugation. Cells were labeled with antibodies against CD3 (T lymphocytes), CD14 (macrophages) and G1 (neutrophils) and cell composition was determined by flow cytometry. A Principal Component Analysis of the cell composition of milk samples was performed on the percent values for each of the three cell types analyzed. Dots are colored depending on the cluster to which they were allocated based on the hierarchical clustering analysis of gene expression data. Color code is identical to that of and is indicated on the figure.

Article Snippet: Antibodies used in the present study were CD14-PE-AF750 (Bio-Rad, reference MCA1568P750, clone Tük4), CD45-PE (Bio-Rad, reference MCA2220PE, clone 1.11.32), G1 (Kingfisher Biotech, reference WSC0608B-100, clone CH138A) and CD3 (Kingfisher Biotech, reference WS0561B-100, clone MM1A).

Techniques: Centrifugation, Labeling, Flow Cytometry, Gene Expression

Journal: Biology

Article Title: Supra-Physiological Levels of Magnesium Counteract the Inhibitory Effect of Zoledronate on RANKL-Dependent Osteoclastogenesis

doi: 10.3390/biology14050533

Figure Lengend Snippet: Flow cytometry analysis of surface antigens in THP-1-derived osteoclasts after a 7-day stimulation with M-CSF/RANKL and parallel treatment with ZA/MgCl 2 . Expression of the CD14 and CD11b surface antigens was assessed by the flow cytometry analysis of THP1 cells differentiated to osteoclasts upon a 2-day incubation with PMA followed by 7-day stimulation with M-CSF and RANKL. Contextually, these cells were also treated with zoledronate (ZA), MgCl 2 (Mg), both compounds (ZA + Mg) or neither of the two (Ctr). Upper panel shows a bar histogram in which the percentage of CD14 positivity is reported on the y-axis and the tested treatments are indicated on the x-axis. Data are presented as the mean ± S.E.M. values of four independent experiments. Lower panel shows the flow cytometry dot plots obtained in a representative experiment after a double labeling of the CD14 and CD11b antigens. The tested treatments are indicated on the top of each dot plot, whereas percentages of cell subpopulations, exhibiting a differential expression of the two analyzed antigens, are reported inside quadrants. Double positive (CD14+CD11b+) cells are represented in red, whereas all other cell subpopulations are represented in grey.

Article Snippet: The entity of differentiation was evaluated after an incubation for 30 min at 4 °C, in PBS containing 5% FCS and 1% FcR blocking reagent (Miltenyi Biotec, Auburn, CA, USA), in the presence of fluorescein isothiocyanate-conjugated (FITC) mouse anti-human CD14 monoclonal antibody (MoAb) and phycoerythrin-conjugated (PE) mouse anti-human CD11b MoAb (Miltenyi Biotec, Auburn, CA, USA), both diluted at a 1:100 ratio.

Techniques: Flow Cytometry, Derivative Assay, Expressing, Incubation, Labeling, Quantitative Proteomics

Journal: Biology

Article Title: Supra-Physiological Levels of Magnesium Counteract the Inhibitory Effect of Zoledronate on RANKL-Dependent Osteoclastogenesis

doi: 10.3390/biology14050533

Figure Lengend Snippet: Flow cytometry analysis of surface antigens in THP-1-derived osteoclasts after a 14-day stimulation with M-CSF/RANKL and a parallel treatment with ZA/MgCl 2 . Expression of CD14 and CD11b surface antigens was assessed by flow cytometry in THP-1 cells differentiated to osteoclasts upon a 2-day incubation with PMA followed by 14-day stimulation with M-CSF and RANKL. Treatment conditions, analysis modalities, and data presentation are the same as in .

Article Snippet: The entity of differentiation was evaluated after an incubation for 30 min at 4 °C, in PBS containing 5% FCS and 1% FcR blocking reagent (Miltenyi Biotec, Auburn, CA, USA), in the presence of fluorescein isothiocyanate-conjugated (FITC) mouse anti-human CD14 monoclonal antibody (MoAb) and phycoerythrin-conjugated (PE) mouse anti-human CD11b MoAb (Miltenyi Biotec, Auburn, CA, USA), both diluted at a 1:100 ratio.

Techniques: Flow Cytometry, Derivative Assay, Expressing, Incubation