rat anti-mouse major histocompatibility complex (mhc) class ii  (Thermo Fisher)

Journal: PLoS ONE

Article Title: Species Difference of CD137 Ligand Signaling in Human and Murine Monocytes

doi: 10.1371/journal.pone.0016129

Figure Lengend Snippet: Monocytes were cultured under indicated conditions for 7 days. (A) Expression of CD80, CD86, F4/80 and CD14 were determined by flow cytometry. Numbers in the graphs represent the percentages of positively stained cells (grey histograms) compared to the isotype control (open histograms). (B) Expression of CD11c and MHC-II were determined by flow cytometry. Numbers in the graphs represent percentage of the population in each quadrant. (C) Cytokine secretion. The concentrations of (A) IL-10, (B) IL-12, and (C) soluble CD137 in the supernatants were measured by ELISA. *p<0.05. N.D: Not detectable. (D) Phagocytosis was determined by adding flourescent beads to cells at a ratio of 50∶1. The flourescence was determined by flow cytometry. Numbers in the graphs indicate the percentages of positive cells and mean fluorescence intensities (MFI). Half of the harvested cells were trypsinized to remove non-phagozytosed beads that might have stuck on the surface of the cells. Data are representative of three independent experiments.

Article Snippet: Phycoerythrin (PE)-conjugated and Flourescein isothiocyanate (FITC) labeled rat anti-mouse mouse CD11c, CD14, CD80, CD86, F4/80, major histocompatibility complex (MHC) class II, and respective isotype controls (rat IgG2a, rat IgG2b, Armenian Hamster IgG) were purchased from eBioscience (San Diego, USA).

Techniques: Cell Culture, Expressing, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: PLoS ONE

Article Title: Species Difference of CD137 Ligand Signaling in Human and Murine Monocytes

doi: 10.1371/journal.pone.0016129

Figure Lengend Snippet: (A) Source of cells: Balb/C CD11b + cells from bone marrow and spleen and total bone marrow cells were treated with immobilized Fc or CD137-Fc protein. (B, C) Source of CD137 protein: CD11b + , Ly6G − monocytes were treated with immobilized Fc or human or murine CD137-Fc protein. Cells differentiated to immature and mature classical DCs were included as controls. (B) After 7 days the cells were used as stimulators in a MLR with C57/Bl6 T cells at a ratio of 1∶10. Proliferation was determined by 3 H-thymidine incorporation at day 3 of coculture. (C) Expression of CD14 and F4/80 was determined at day 7 of the culture. iDC: immature DC. *p<0.05. Data are representative of three independent experiments.

Article Snippet: Phycoerythrin (PE)-conjugated and Flourescein isothiocyanate (FITC) labeled rat anti-mouse mouse CD11c, CD14, CD80, CD86, F4/80, major histocompatibility complex (MHC) class II, and respective isotype controls (rat IgG2a, rat IgG2b, Armenian Hamster IgG) were purchased from eBioscience (San Diego, USA).

Techniques: Expressing

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: High-mobility group box 1 is an important mediator of microglial activation induced by cortical spreading depression

doi: 10.1177/0271678X16647398

Figure Lengend Snippet: Multiple CSD-induced microglial activation in the cerebral cortex is dependent on HMGB1 activity. (a) The total number of Iba1-positive microglia in the cerebral cortex. n = 6 in each group. (b) Immunostaining of the cerebral cortex for Iba1 in the control, sham, CSD1x-24 h, and CSD5x-24 h groups (the upper row). Nuclear staining was performed with DAPI (the middle row). Merged images are shown in the lower row. Bar: 10 µm. (c) The number of enlarged Iba1-positive microglia. (d) The proportion of microglial somal area to the entire brain tissue area. Values are presented as mean ± SD. Statistical analysis was carried out by ANOVA and Bonferroni’s post hoc test.

Article Snippet: We also conducted immunostaining employing mouse antisera against major histocompatibility complex (MHC) class II antigen (1:500; MCA46GA; AbD Serotec, Raleigh, NC, USA) and goat antisera against cathepsin D (1:500; sc-6486; Santa Cruz Biotechnology, Santa Cruz, Dallas, TX, USA).

Techniques: Activation Assay, Activity Assay, Immunostaining, Control, Staining

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: High-mobility group box 1 is an important mediator of microglial activation induced by cortical spreading depression

doi: 10.1177/0271678X16647398

Figure Lengend Snippet: Upregulation of cathepsin D expression in hypertrophic microglia induced by multiple CSDs. Representative microglia immunostained for Iba1 (green) and cathepsin D (red) in the cerebral cortex of the control and CSD5x-24 h group mice. Granular immunostaining patterns of cathepsin D, consistent with intracellular lysosomal localization, are indicated by arrowheads. Bar: 10 µm.

Article Snippet: We also conducted immunostaining employing mouse antisera against major histocompatibility complex (MHC) class II antigen (1:500; MCA46GA; AbD Serotec, Raleigh, NC, USA) and goat antisera against cathepsin D (1:500; sc-6486; Santa Cruz Biotechnology, Santa Cruz, Dallas, TX, USA).

Techniques: Expressing, Control, Immunostaining

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: High-mobility group box 1 is an important mediator of microglial activation induced by cortical spreading depression

doi: 10.1177/0271678X16647398

Figure Lengend Snippet: (a) Representative immunostaining for Iba1 and nuclear counterstaining in the TLR2/4 KO mouse cerebral cortex. Bar: 10 µm. The numbers of total (b) and enlarged (c) Iba1-positive microglia in the cerebral cortex of the wild-type and TLR2/4 KO mice in the control and CSD5x-24 h groups. (d) The proportion of microglial somal area to the entire brain tissue area. Values are expressed as mean ± SD. Statistical analysis was carried out by ANOVA and Bonferroni’s post hoc test.

Article Snippet: We also conducted immunostaining employing mouse antisera against major histocompatibility complex (MHC) class II antigen (1:500; MCA46GA; AbD Serotec, Raleigh, NC, USA) and goat antisera against cathepsin D (1:500; sc-6486; Santa Cruz Biotechnology, Santa Cruz, Dallas, TX, USA).

Techniques: Immunostaining, Control