biotin conjugated anti human il 10  (R&D Systems)

Journal: Veterinary Research

Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

doi: 10.1186/1297-9716-44-59

Figure Lengend Snippet: Effect of PD-1 blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P < 0.05 (* P < 0.05; ** P < 0.01).

Article Snippet: Sandwich ELISA of IL-10 was performed with two antibodies; anti-IL-10 (CC318; AbD Serotec) as capture antibody and biotin-conjugated anti-IL-10 (CC320; AbD Serotec) as the detective antibody.

Techniques: Cell Function Assay, Cell Culture, Negative Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Staining

Journal: Obesity (Silver Spring, Md.)

Article Title: Tight Association between Macrophages and Adipocytes in Obesity: Implications for Adipocyte Preparation

doi: 10.1002/oby.20634

Figure Lengend Snippet: Most CLS macrophages express TNFα while a few express both TNFα and IL-10. Images presented are a LSCM 2D projection of a 3D image stack. Adipose tissue from obese mice that was fixed and stained with BODIPY 558/568 for lipid (gray), Hoechst for nuclei (blue), anti-F4/80 for macrophages (red), anti-TNFα (green), and anti-IL-10 (magenta). The panel shows a four micron 2D projection captured with a 63× objective. The majority of CLS macrophages produce TNFα and some appear to produce both TNFα and IL-10 (arrows) while others have no cytokine expression (F4/80 only).

Article Snippet: For simple staining, fixed adipose tissue and floating layer cells were stained with rat anti-mouse F4/80 (Invitrogen, Carlsbad, CA) to mark macrophages and detected with donkey anti-rat IgG conjugated to AlexaFluor488 (Invitrogen) or IL-10 was stained with anti-mouse IL-10 biotin conjugate (Invitrogen) and an Alexa647 conjugated streptavidin (Invitrogen).

Techniques: Staining, Expressing

Journal: Obesity (Silver Spring, Md.)

Article Title: Tight Association between Macrophages and Adipocytes in Obesity: Implications for Adipocyte Preparation

doi: 10.1002/oby.20634

Figure Lengend Snippet: The adipocyte floating layer has high expression of TNFα and IL-10 in obese mice compared to non-obese mice. (a) qRT-PCR of floating layer cells showing relative amounts of TNFα compared to IL-10 mRNA normalized to GAPDH. Obese mice had a 40-fold increase in TNFα (open boxes) while only inducing a 10-fold increase in IL-10 production (black boxes). * p<0.05 in all figures. (b) Floating layer adipocyte with attached macrophages. Floating layer cells stained with BODIPY (gray) for lipid to denote adipocytes, F4/80 (red) for macrophages, TNFα (green), IL-10 (magenta), and nuclei (blue). There was no IL-10 expressed in either cell type in this example (field not shown). TNFα can clearly be seen to be associated with only the macrophages, but not the adipocyte (F4/80 vs. TNFα panels and overlay). The adipocyte nucleus is indicated by the white arrow in the overlay and right side view, while macrophage nuclei are indicated by the blue arrows. The figure is an LSCM 2D projection of a 3D image stack using a 63× objective. The side views are 1um thick sections imaged from the corresponding dotted white lines parallel to the section.(c) Macrophages in the floating adipocyte layer ingest lipid and produce TNFα. At high magnification using LSCM and 63× objective a 2D projection shows a macrophage (red) with a single nucleus (blue arrow) remained attached to and ingested lipid (gray) in the floating layer. TNFα (green) expression was associated with the macrophage (white arrows) and not the attached or ingested lipid. No IL-10 is produced. (d) Floating layer macrophage(s) with engulfed lipid produced IL-10 to a lesser extent. Individual panels (left) show a one micron section of a LSCM image using a 63× objective. The floating layer cells were stained for F4/80 (red), TNFα (green) and IL-10 (magenta) and the overlay depicts all of these fields plus a brightfield image. The macrophage, shown with engulfed lipid, is expressing mostly IL-10, but there is some expression of TNFα. A 2D projection of the LSCM image shows the extent of IL-10 production in the macrophage surrounding the lipid.

Article Snippet: For simple staining, fixed adipose tissue and floating layer cells were stained with rat anti-mouse F4/80 (Invitrogen, Carlsbad, CA) to mark macrophages and detected with donkey anti-rat IgG conjugated to AlexaFluor488 (Invitrogen) or IL-10 was stained with anti-mouse IL-10 biotin conjugate (Invitrogen) and an Alexa647 conjugated streptavidin (Invitrogen).

Techniques: Expressing, Quantitative RT-PCR, Staining, Produced