agonist anti trem 1 monoclonal antibody  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: GM1 Oligosaccharide Modulates Microglial Activation and α-Synuclein Clearance in a Human In Vitro Model

doi: 10.3390/ijms26157634

Figure Lengend Snippet: Effects of GM1 oligosaccharide (OligoGM1) application on Human embryonic microglial (HCM3) cells. After 2 days of culture (80% of confluence), HCM3 cells were incubated or not incubated (control, CTRL) with OligoGM1 (100 µM). After 48 h, immunofluorescence staining [(against nuclei, ionized calcium-binding adaptor molecule 1 (Iba1) and triggered receptor expressed on myeloid cells 2 (TREM2)] and ELISA for quantification of released cytokines to medium were performed, as described in the : ( a ) Representative immunofluorescence images (20× magnification, scale bar 100 µm) and number of DAPI-positive cells; ( b ) Representative immunofluorescence images (20× magnification, scale bar 100 µm) of Iba1 expression, quantification of Iba1 accumulation (Iba1 area normalized by the number of cells), and area of Iba1-positive cells (area of microglia cells normalized by the µm 2 of Iba1); ( c ) Representative immunofluorescence images (20× magnification, scale bar 100 µm) of TREM2 expression, quantification of TREM2-positive cells (TREM2 area normalized by the number of cells), and area of TREM2-positive cells (area of microglia cells normalized by the µm 2 of TREM2); ( d ) Quantification of released cytokines tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6). All values are represented as percentage versus CTRL and expressed as mean ± standard error of the mean (SEM, n = 6; ns = not significant, Mann–Whitney test).

Article Snippet: For immunofluorescence analyses, the following antibodies were used: primary mouse monoclonal anti-TREM2 antibody [Cat. 11084-MM08, research resource identifier (RRID):AB_2860315], purchased from Sino Biological Europe GmbH, Europe (Düsseldorfer, Germany); primary rabbit polyclonal anti-αSyn antibody (Cat. 2642S, RRID not available), purchased from Ozyme, Paris, France; and primary goat polyclonal anti-Iba1 antibody (Cat. Ab5076, RRID:AB_2224402), purchased from Abcam, Cambridge, UK.

Techniques: Incubation, Control, Immunofluorescence, Staining, Binding Assay, Enzyme-linked Immunosorbent Assay, Expressing, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: GM1 Oligosaccharide Modulates Microglial Activation and α-Synuclein Clearance in a Human In Vitro Model

doi: 10.3390/ijms26157634

Figure Lengend Snippet: OligoGM1 modulation of αSyn-induced activation of HMC3 cells. After 2 days of culture (80% of confluence), HCM3 cells were incubated or not incubated (CTRL) with OligoGM1 (100 µM). After 4 h, αSyn (1 µM) was added to the culture medium for 48 h. At the end of treatment, immunofluorescence staining (against nuclei, Iba1, TREM2, and αSyn) and IL-6/TNF-α quantification from medium were performed as described in the . ( a ) Representative immunofluorescence images (20× magnification, scale bar 100 µm) and number of DAPI-positive cells; ( b ) Representative immunofluorescence images (20× magnification, scale bar 50 µm) of Iba1 expression, quantification of Iba1 accumulation (Iba1 area normalized by the number of cells), and area of Iba1-positive cells (area of microglia cells normalized by the µm 2 of Iba1); ( c ) Representative immunofluorescence images (20× magnification, scale bar 25 µm) of TREM2 and αSyn expressions, quantification of TREM2-positive cells (TREM2 area normalized by the number of cells), area of TREM2-positive cells (area of microglia cells normalized by the µm 2 of TREM2), and quantification of αSyn area accumulated within TREM(+) cells (area of αSyn normalized by the µm 2 of TREM2); ( d ) Quantification of released cytokines TNF-α and IL-6. All values are represented as percentage versus CTRL and expressed as mean ± SEM ( n = 6; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = not significant by one-way ANOVA followed by Fisher’s LSD). * p < 0.05 was considered significant.

Article Snippet: For immunofluorescence analyses, the following antibodies were used: primary mouse monoclonal anti-TREM2 antibody [Cat. 11084-MM08, research resource identifier (RRID):AB_2860315], purchased from Sino Biological Europe GmbH, Europe (Düsseldorfer, Germany); primary rabbit polyclonal anti-αSyn antibody (Cat. 2642S, RRID not available), purchased from Ozyme, Paris, France; and primary goat polyclonal anti-Iba1 antibody (Cat. Ab5076, RRID:AB_2224402), purchased from Abcam, Cambridge, UK.

Techniques: Activation Assay, Incubation, Immunofluorescence, Staining, Expressing

Journal: EMBO Molecular Medicine

Article Title: Female sex is linked to a stronger association between sTREM2 and CSF p-tau in Alzheimer’s disease

doi: 10.1038/s44321-024-00190-3

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Monoclonal mouse anti-human TREM2 antibody , Santa Cruz Biotechnology; B-3, sc373828.

Techniques: Recombinant, Sequencing, Software

Journal: International Journal of Molecular Sciences

Article Title: The Interaction of HMGB1 with the Proinflammatory TREM-1 Receptor Generates Cytotoxic Lymphocytes Active against HLA-Negative Tumor Cells

doi: 10.3390/ijms25010627

Figure Lengend Snippet: ( a ) The HMGB1 protein was applied to sTREM-1 conjugated with Sepharose, eluted with acetonitrile, and resolved using SDS PAGE and Western blot with antibodies to HMGB1. ( b ) ELISA of soluble TREM-1 in lymphocytes in presence of different concentrations of HMGB1 protein. The cultured medium was collected after 24 h of incubation with various concentrations of HMGB1. Experiment was carried out in triplicate, and the most common data are shown.

Article Snippet: The TREM-1 receptor was stained with monoclonal mouse antibodies against TREM-1 and Goat anti-mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (Molecular Probes By Life Technologies, Carlsbad, CA, USA), and for HMGB1–polyclonal rabbit antibodies against HMGB1 and Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 633 (Molecular Probes By Life Technologies, Carlsbad, CA, USA).

Techniques: SDS Page, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, Incubation

Journal: International Journal of Molecular Sciences

Article Title: The Interaction of HMGB1 with the Proinflammatory TREM-1 Receptor Generates Cytotoxic Lymphocytes Active against HLA-Negative Tumor Cells

doi: 10.3390/ijms25010627

Figure Lengend Snippet: Confocal micrograph of HMGB1 (green) and TREM-1 (red) and layers superposition on the surface of monocyte. Arrows indicate colocalization region. About 50% of all observed cells were double stained. (n = 4) For analysis ImageJ Software (V 1.8.0) was used.

Article Snippet: The TREM-1 receptor was stained with monoclonal mouse antibodies against TREM-1 and Goat anti-mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (Molecular Probes By Life Technologies, Carlsbad, CA, USA), and for HMGB1–polyclonal rabbit antibodies against HMGB1 and Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 633 (Molecular Probes By Life Technologies, Carlsbad, CA, USA).

Techniques: Staining, Software