recombinant human his tagged trem2  (Sino Biological)

Journal: bioRxiv

Article Title: Identification of cellular and molecular risk signatures for progression to late-stage age-related macular degeneration using the 9-step Minnesota Grading System

doi: 10.1101/2025.07.10.664209

Figure Lengend Snippet: A . Flowchart describing selection of gene markers associated with a high-risk of progression to late AMD for validation, based on bulk RNAseq of the macular RPE/choroid. B. A heatmap showing high-risk marker gene expression in the macular RPE/choroid in samples of AMD1, AMD3L, and AMD3H. The levels of each mRNA are expressed as log 2 (TPM+1), where TPM stands for transcript per million, red indicates positive value and blue negative. C . Representative RNAscope images of C3 mRNA in AMD1, AMD3L, AMD3H, and AMD4/GA macula sections, with arrows indicating C3 mRNA + signals. C3 increases in the RPE-adjacent region with disease severity. D. Representative RNAscope images of TREM2 mRNA in AMD1, AMD3L, AMD3H, and AMD4/GA macula sections, with arrows indicating TREM2 mRNA+ signals. TREM2 mRNA becomes apparent in the choroid at AMD3H and persists in AMD4. E. ELISA measurement of TREM2 protein in AMD1 (n = 8 samples) and AMD4 (n = 6-8) in serum, macular and peripheral RPE/choroid tissue and neuroretina, vitreous, and aqueous humor. Donor information is in . The pM label indicates picomolar concentrations, with p value determined by unpaired Student’s t-test (* p < 0.05, *** P <0.001). F . Representative RNAscope images of OLR1 mRNA in AMD1, AMD3L, AMD3H, and AMD4/GA macula sections, with arrows indicating OLR1 mRNA+ signals. OLR1 increases in the RPE-adjacent region with disease severity. G . Representative RNAscope images of NGFR mRNA in AMD1, AMD3L, AMD3H, and AMD4/GA macula sections, with arrows indicating NGFR mRNA+ signals. Elevated NGFR signal is observed in AMD3H, proximal to the RPE.

Article Snippet: Total TREM2 protein levels were measured using a TREM2 Duoset (R&D Systems, Minneapolis, MN) ELISA kit for MSD-based assays (MSD, Rockville, MD) and read on a Sector S 600 imager.

Techniques: Selection, Biomarker Discovery, Marker, Gene Expression, RNAscope, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Identification of cellular and molecular risk signatures for progression to late-stage age-related macular degeneration using the 9-step Minnesota Grading System

doi: 10.1101/2025.07.10.664209

Figure Lengend Snippet: Fibroblasts predominantly express C3, immune cells show C3, OLR1 and TREM2 expression, and GFAP, NGFR, SPP1 and GPX2 are expressed in Muller glia and astrocytes.

Article Snippet: Total TREM2 protein levels were measured using a TREM2 Duoset (R&D Systems, Minneapolis, MN) ELISA kit for MSD-based assays (MSD, Rockville, MD) and read on a Sector S 600 imager.

Techniques: Expressing

Journal: bioRxiv

Article Title: Identification of cellular and molecular risk signatures for progression to late-stage age-related macular degeneration using the 9-step Minnesota Grading System

doi: 10.1101/2025.07.10.664209

Figure Lengend Snippet: A . A dot plot of TREM2 and OLR1 expression in all detected RPE/choroid cell types. Both TREM2 and OLR2 are preferentially expressed in choroidal macrophages. Of the three clusters of macrophages, TREM2 is expressed in macrophage 1 and 3 while OLR1 is expressed in macrophage 2 and 3. B . Dual genes plot of all 3 macrophage populations expressing TREM2 (green dots) versus OLR1 (red dots). There are very few macrophages (yellow dots) expressing both TREM2 and OLR1 .

Article Snippet: Total TREM2 protein levels were measured using a TREM2 Duoset (R&D Systems, Minneapolis, MN) ELISA kit for MSD-based assays (MSD, Rockville, MD) and read on a Sector S 600 imager.

Techniques: Expressing

Journal: bioRxiv

Article Title: Identification of cellular and molecular risk signatures for progression to late-stage age-related macular degeneration using the 9-step Minnesota Grading System

doi: 10.1101/2025.07.10.664209

Figure Lengend Snippet: In AMD3H, complement C3 can be induced in multiple neuronal and choroidal cell-types including fibroblasts, macrophages, and Müller/astrocytes. Macrophages expressing either TREM2 or OLR1 are observed near the RPE/choriocapillaris complex in AMD3H, possibly originating from the choroid and/or from the systemic circulation, and later in AMD3H/4 outer retina. Activated Müller/astrocytes cells expressing NGFR and GFAP expand from the inner retina to outer retina and RPE in AMD3H. Activation of complement, macrophages, and Müller/astrocytes may signal the onset of the end stage AMD. Graphic was created with Biorender ( biorender.com ).

Article Snippet: Total TREM2 protein levels were measured using a TREM2 Duoset (R&D Systems, Minneapolis, MN) ELISA kit for MSD-based assays (MSD, Rockville, MD) and read on a Sector S 600 imager.

Techniques: Expressing, Activation Assay

Journal: bioRxiv

Article Title: Identification of cellular and molecular risk signatures for progression to late-stage age-related macular degeneration using the 9-step Minnesota Grading System

doi: 10.1101/2025.07.10.664209

Figure Lengend Snippet:

Article Snippet: Total TREM2 protein levels were measured using a TREM2 Duoset (R&D Systems, Minneapolis, MN) ELISA kit for MSD-based assays (MSD, Rockville, MD) and read on a Sector S 600 imager.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of neuroinflammation

Article Title: Accumulated BCAAs and BCKAs contribute to the HFD-induced deterioration of Alzheimer's disease via a dysfunctional TREM2-related reduction in microglial β-amyloid clearance.

doi: 10.1186/s12974-024-03314-1

Figure Lengend Snippet: Fig. 4 BCAAs and BCKAs impact the response of microglial TREM2 and autophagy to Aβ. A Western blot analysis of TREM2-related protein levels in cortex of APP/PS1 mice and the corresponding quantification results (n = 4). B Relative mRNA levels of DAP12 and CD68 in cortex (n = 5). C Western blot analysis of autophagy-related protein levels in cortex and the corresponding quantification results (n = 4). D Relative mRNA levels of BECLIN1 and LAMP1 in cortex (n = 5). Western blot analysis of TREM2-related E and autophagy-related F Protein levels in BV2 cell treated with or without BCAAs/ BCKAs for 4 h in the presence of Aβ and the corresponding quantification results (n = 3). Western blot analysis of TREM2-related G and autophagy-related H protein levels in BV2 cell treated with or without BCAAs/ BCKAs for 12 h in the presence of Aβ and the corresponding quantification results (n = 3). Data are means ± SEM. *P < 0.05,**P < 0.01 and ***P < 0.001, one-way ANOVA, followed by Tukey’s multiple comparisons test

Article Snippet: The recombinant human TREM2 protein was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Western Blot

Journal: Journal of neuroinflammation

Article Title: Accumulated BCAAs and BCKAs contribute to the HFD-induced deterioration of Alzheimer's disease via a dysfunctional TREM2-related reduction in microglial β-amyloid clearance.

doi: 10.1186/s12974-024-03314-1

Figure Lengend Snippet: Fig. 5 BCAAs and BCKAs suppress TREM2 activation through binding to TREM2. A Molecular docking results of L-Leucine and KICA binding to TREM2 extracellular domain. B Biolayer interferometry (BLI) analysis of interaction of BCAAs or BCKAs with TREM2. BCAAs and BCKAs was titrated from 15.63 to 500 μM and 31.25 to 500 μM, respectively. C LC–MS/MS spectra of TREM2 immunoprecipitated L-Leucine-13C of BV2 cells after BCAAs isotope treatment for 1 h. D Thermal stabilization of TREM2 protein evaluated by cellular thermal shift assay (CETSA) after treating BV2 cells with BCAAs/ BCKAs for 4 h (n = 3). E Representative images of binding of fluorescein labeled-Aβ to cell surface TREM2 in BV2 cells after treatment of BCAAs/ BCKAs for 2 h (n = 3, Scale bars, 40 μm). F Representative images of microglial phagocytosis of fluorescein labeled-Aβ in BV2 cells after treatment of BCAAs/ BCKAs in the presence or absence of S1P for 4 h (n = 3, Scale bars, 40 μm). G Western blot analysis of phosphorylation levels of SYK in BV2 cell after treatment of BCAAs/ BCKAs in the presence or absence of sphingosine-1-phosphate (S1P) for 4 h (n = 3). Data in D, E, F and G are means ± SEM. *P < 0.05,**P < 0.01 and ***P < 0.001, one-way ANOVA, followed by Tukey’s multiple comparisons test

Article Snippet: The recombinant human TREM2 protein was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Immunoprecipitation, Thermal Shift Assay, Labeling, Western Blot, Phospho-proteomics

Journal: Journal of neuroinflammation

Article Title: Accumulated BCAAs and BCKAs contribute to the HFD-induced deterioration of Alzheimer's disease via a dysfunctional TREM2-related reduction in microglial β-amyloid clearance.

doi: 10.1186/s12974-024-03314-1

Figure Lengend Snippet: Fig. 6 BCAAs and BCKAs reduce the expression and recycling of microglial TREM2 via inhibiting autophagy. Western blot analysis of TREM2-related A and autophagy-related B protein levels in BV2 cell after treatment of BCAAs/ BCKAs in the presence or absence of rapamycin for 12 h (n = 3). C Representative images of microglial phagocytosis of fluorescein labeled-Aβ in BV2 cells after treatment of BCAAs/ BCKAs in the presence or absence of rapamycin for 12 h (n = 3, Scale bars, 40 μm). D Representative images of TREM2 recycling in BV2 cells after treatment of BCAAs/ BCKAs in the presence or absence of rapamycin for 12 h (n = 3, Scale bars, 40 μm). Western blot analysis of TREM2-related E and autophagy-related F protein levels in BV2 cell after treatment of BCAAs/ BCKAs in the presence or absence of chloroquine for 12 h (n = 3). G Representative images of TREM2 recycling in BV2 cells after treatment of BCAAs/ BCKAs in the presence or absence of chloroquine for 12 h (n = 3, Scale bars, 40 μm). Data are means ± SEM. *P < 0.05,**P < 0.01 and ***P < 0.001, one-way ANOVA, followed by Tukey’s multiple comparisons test or Student’s t test

Article Snippet: The recombinant human TREM2 protein was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, Labeling