cd14 af700 antibody  (Thermo Fisher)

Journal: Veterinary immunology and immunopathology

Article Title: Flow cytometry analysis of CD11c-positive peripheral blood mononuclear cells in horses.

doi: 10.1016/j.vetimm.2022.110504

Figure Lengend Snippet: Fig. 2. Cell immunophenotyping of horse PBMCs using a panel with CD11c, MHC-II, CD14 and TLR4 and cross-reactivity assays. (a) Gate strategy to remove doublets in the forward and side scatter with height to area and cell population identification of population 1 (P1), population 2 (P2) and population 3 (P3). (b) Cell expression dot plot of CD11c+CD14+, CD11c+MHCII+ and CD11c+TLR4+ P1, P2 and P3. (c) Cross-reactivity assay of antibodies for their recognition capacity in equine PBMCs. Black histograms represent the negative control, and colored histograms are representative of P1, P2 and P3. Data representative from one horse. Samples were analyzed by flow cytometry; 50,000 events were obtained. The analysis was performed using FlowJo V10 software.

Article Snippet: The cells were incubated with other surface markers using the following antibodies: anti-horse MHCII-FITC (Batch No. 0515, Bio-Rad), CD14-AF700 (FAB4597N, Novus Biologicals), and TLR4-PE (clone: HTA125, Cat: 312805, Bio-Legend).

Techniques: Expressing, Negative Control, Flow Cytometry, Software

Journal: Journal of Medicinal Chemistry

Article Title: A Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Assay Identifies Nilotinib as an Inhibitor of Inflammation in Acute Myeloid Leukemia

doi: 10.1021/acs.jmedchem.2c00671

Figure Lengend Snippet: Nilotinib inhibits cell adhesion and monocyte activation. (A) PCA plot of log 2-transformed LFQ intensities from THP-1 cells pre-treated with DMSO, 5 μM nilotinib, or imatinib for 1 h before stimulation with live E. coli for up to 24 h, showing distinct grouping of treatments. (B,C) Volcano plot of THP-1 cells treated with (B) nilotinib vs imatinib and (C) nilotinib vs E. coli , a cutoff of FDR <0.05, and a 1.5-fold change between conditions. (D) Expression levels of CD44, (E) CD11c, (F) CD14, (G) CD49a, and (H) CD54 on the cell surface were measured by flow cytometry. (I) Optical density of dissolved crystal violet was used to evaluate the adhesion rate. Significant differences between two groups were determined by the Mann–Whitney U-test. The statistical significance of the comparisons with E. coli is indicated as follows: ns, not significant; *, P ≤ 0.05; **, P ≤ 0.01. Error bars represent the standard deviation of four biological replicates.

Article Snippet: Cell surface staining was performed by the direct immunofluorescence assay with fluorescent-conjugated antibodies: CD11c-APC (#17-0114-82), CD14-AF700 (#56-0149-42), and CD54-FITC (#11-0541-82) from Thermo Fisher; CD49a-PE (#562115) and CD44-FITC (#338803) from BD Biosciences; and corresponding isotype control antibodies, for 30 min at 4 °C in PBS with 1% FBS, 1% BSA, and 1% human serum (Sigma) to block Fc receptors.

Techniques: Activation Assay, Transformation Assay, Expressing, Flow Cytometry, MANN-WHITNEY, Standard Deviation

Journal: Cell reports

Article Title: Single-Cell Analysis Suggests that Ongoing Affinity Maturation Drives the Emergence of Pemphigus Vulgaris Autoimmune Disease

doi: 10.1016/j.celrep.2019.06.066

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD14-AF700 (61D3) , eBioscience , Cat# 56-0149-42; RRID: AB_2574497.

Techniques: Virus, Recombinant, Avidin-Biotin Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, DNA Purification, Transfection, Modification, Sequencing, Cloning, Expressing

Journal: Theranostics

Article Title: Glyco-Dendrimers as Intradermal Anti-Tumor Vaccine Targeting Multiple Skin DC Subsets

doi: 10.7150/thno.35059

Figure Lengend Snippet: Enhanced glyco-dendrimer binding and uptake by moDC via DC-SIGN and primary LC via Langerin. Binding and uptake of G3 (glyco)-dendrimers was evaluated for DC-SIGN + moDC and Langerin + primary LCs. A. Dose-response following a 3 hour pulse, wash and 45 minutes chase of moDC with G3 (glyco)-dendrimers in the presence (dotted line) or absence (solid line) of TLR4 stimulus MPLA. Representative of n=3 measured in triplicate ±SD B. Binding and uptake of (glyco)-dendrimers over-time by moDC following a 45 minutes pulse (no wash) at 4 o C. n=2, ±SD C. Imaging microscopy of moDC following 3 hour incubation at 37 o C with glyco-dendrimers (green). Membrane was stained using anti-CD1a (red) and nucleus using DAPI (white) D-E. Involvement of DC-SIGN in binding and uptake of G3 (glyco)- dendrimers was evaluated using a 3 hour pre-incubation with anti-DC-SIGN (C) or 30 minutes pre-incubation with the natural ligand mannan (D) followed by 1 hour incubation with (glyco)-dendrimers. (C) n=4, each symbol represents a donor, (D) representative of n=2 measured in triplicate ±SD F . Binding and uptake of (glyco)-dendrimers over-time by primary LC following a 45 minutes pulse (no wash) on 4 o C. Representative of n=2 measured in triplicate G. Langerin involvement in binding and uptake of (glyco)-dendrimers by primary LCs was evaluated using 30 minutes pre-incubation an anti-Langerin blocking antibody followed by 1 hour co-incubation with (glyco)-dendrimers. Representative of n=3 measured in triplicate ±SD H. Imaging microscopy of primary LC following 3 hours incubation at 37 o C with glyco-dendrimers (green). Membrane was stained using anti-CD1a (red) (Statistical analysis: A,E two-way ANOVA Sidak's post hoc; B,D,F two-way ANOVA Tukey's post hoc; C one-way ANOVA Dunnett's post hoc)

Article Snippet: To distinguish the different emigrated skin DC subsets cells were stained using the following anti-human antibodies: HLA-DR BV510, CD1a APC, CD14 AF700 (clone M5E2, Sony), CD141 BV711 (clone 1A4, Biolegend), EpCAM BV421 and FVD.

Techniques: Binding Assay, Imaging, Microscopy, Incubation, Membrane, Staining, Blocking Assay