mouse anti trem2 primary antibody  (Proteintech)

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: Frontiers in Neuroscience

Article Title: Perioperative enriched environment attenuates postoperative cognitive dysfunction by upregulating microglia TREM2 via PI3K/Akt pathway in mouse model of ischemic stroke

doi: 10.3389/fnins.2024.1520710

Figure Lengend Snippet: EE upregulated the expression of TREM2 in the hippocampus after surgery in mice with ischemic stroke. (A,B) The expression of TREM2 protein in the hippocampus on day 10 after surgery ( n = 3). (C,D) The expression of DAP12 protein in the hippocampus on day 10 after surgery ( n = 3). (E) The expression of TREM2 mRNA in the hippocampus on day 10 after surgery ( n = 3). (F) The expression of DAP12 mRNA in the hippocampus on day 10 after surgery ( n = 3). (G) The immunofluorescence of the cellular localization of TREM2 in the hippocampus of mice in the Sham+EE group on day 10 after surgery ( n = 3). DG: dentate gyrus. Scale bars represented 10 μm and 50 μm. ns: no significant; * p < 0.05, *** p < 0.001, the Sham+SE group vs. the PT + SE group; # p < 0.05, ## p < 0.01, the PT + SE group vs. the PT + EE group; ^ p < 0.05, ^^^ p < 0.001, the Sham+SE group vs. the Sham+EE group. Error bars were represented as mean ± SD.

Article Snippet: After antigen retrieval with EDTA, the paraffin sections were washed with phosphate-buffered solution (PBS) and then blocked with 5% bovine serum albumin at room temperature for 1 h. Following this, the sections were incubated with a mouse anti-TREM2 primary antibody (1:1,000, 68723-1-Ig, Proteintech) overnight at 4°C.

Techniques: Expressing, Immunofluorescence

Journal: Frontiers in Neuroscience

Article Title: Perioperative enriched environment attenuates postoperative cognitive dysfunction by upregulating microglia TREM2 via PI3K/Akt pathway in mouse model of ischemic stroke

doi: 10.3389/fnins.2024.1520710

Figure Lengend Snippet: TREM2 shRNA successfully decreased the expression of TREM2 in the hippocampus on day 21 after the injection of TREM2 shRNA. (A,B) The expression of TREM2 protein in the hippocampus ( n = 3). (C,D) The expression of DAP12 protein in the hippocampus ( n = 3). (E) The expression of TREM2 mRNA in the hippocampus ( n = 3). (F) The expression of DAP12 mRNA in the hippocampus. ns: no significant; * p < 0.05, *** p < 0.001, the SE + vehicle group vs. the SE + shRNA group; ## p < 0.01, ### p < 0.001, the EE+ vehicle group vs. the EE+ shRNA group. Error bars were represented as mean ± SD.

Article Snippet: After antigen retrieval with EDTA, the paraffin sections were washed with phosphate-buffered solution (PBS) and then blocked with 5% bovine serum albumin at room temperature for 1 h. Following this, the sections were incubated with a mouse anti-TREM2 primary antibody (1:1,000, 68723-1-Ig, Proteintech) overnight at 4°C.

Techniques: shRNA, Expressing, Injection

Journal: Frontiers in Neuroscience

Article Title: Perioperative enriched environment attenuates postoperative cognitive dysfunction by upregulating microglia TREM2 via PI3K/Akt pathway in mouse model of ischemic stroke

doi: 10.3389/fnins.2024.1520710

Figure Lengend Snippet: The knockdown of TREM2 abolished the neuroprotective effects of EE and inhibited the PI3K/Akt signaling pathway in the hippocampus of mice with ischemic stroke after surgery. (A–C) The behavioral tests of each group ( n = 6). (D) Western bolt results of p-PI3K and PI3K. (E) The protein p-PI3K / PI3K ratio in the hippocampus on day 10 after surgery ( n = 3). (F) Western bolt results of p-AKT and AKT. (G) The protein p-AKT/AKT ratio in the hippocampus on day 10 after surgery ( n = 3). ns: no significant; * p < 0.05, *** p < 0.001, the SE + vehicle group vs. the SE + shRNA group; ## p < 0.01, #### p < 0.0001, the EE+ vehicle group vs. the EE+ shRNA group. Error bars were represented as mean ± SD.

Article Snippet: After antigen retrieval with EDTA, the paraffin sections were washed with phosphate-buffered solution (PBS) and then blocked with 5% bovine serum albumin at room temperature for 1 h. Following this, the sections were incubated with a mouse anti-TREM2 primary antibody (1:1,000, 68723-1-Ig, Proteintech) overnight at 4°C.

Techniques: Knockdown, Western Blot, shRNA