Mouse TREM2 Antibody

TREM2 in RAW 264.7 Mouse Cell Line. TREM2 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Sheep Anti-Mouse TREM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1729) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Mouse TREM2 ELISA Standard Curve. Recombinant Mouse TREM2 protein was serially diluted 2-fold and captured by Sheep Anti-Mouse TREM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1729) coated on a Clear Polystyrene Microplate (DY990). Sheep Anti-Mouse TREM2 Biotinylated Antigen Affinity-purified Polyclonal Antibody (BAF1729) was incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

TREM2 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. TREM2 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line (left panel) but is not detected in TREM2 knockout (KO) RAW 264.7 Mouse Cell Line cell line (right panel) using Sheep Anti-Mouse TREM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1729) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Mouse TREM2 by Immunocytochemistry/Immunofluorescence SCF+G-CSF treatment increases TREM2 expression in the Iba1+ microglia/macrophages surrounding the 6E10+ senile plaques. (A) Representative confocal images of TREM2 (red), 6E10 (purple) and Iba1 (green) triple immunofluorescence staining in the brains of aged APP/PS1 mice. Blue: Nuclear counterstaining by DAPI. (B) Representative orthographic view of z-stack images (12 z-stacks with 1μm intervals) illustrates the location and interaction of TREM2 + cells (red) and 6E10+ A beta plaques (white) in the brains of aged APP/PS1 mice. (C) Quantification data show the percentage of TREM2+ area surrounding the 6E10+ A beta plaques (within 10μm from the border of the A beta plaques) in the brains of aged APP/PS1 mice with or without SCF+G-CSF treatment. (D) Representative orthographic view of z-stack images (12 z-stacks with 1μm intervals) displays the location and interaction of TREM2+/Iba1+ co-expressing cells (yellow) and 6E10+ A beta plaques (white) in the brains of APP/PS1 mice. (E) Quantification data show the percentage of TREM2+/Iba1+ co-expression area in the total of Iba1+ area in the vicinity of 6E10+ A beta plaques in the brains of aged APP/PS1 mice with or without SCF+G-CSF treatment. N=4-5. Mean ± SEM. * p<0.05 by Student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33269098), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse TREM2 by Immunocytochemistry/Immunofluorescence SCF+G-CSF treatment increases TREM2 expression in the Iba1+ microglia/macrophages surrounding the 6E10+ senile plaques. (A) Representative confocal images of TREM2 (red), 6E10 (purple) and Iba1 (green) triple immunofluorescence staining in the brains of aged APP/PS1 mice. Blue: Nuclear counterstaining by DAPI. (B) Representative orthographic view of z-stack images (12 z-stacks with 1μm intervals) illustrates the location and interaction of TREM2 + cells (red) and 6E10+ A beta plaques (white) in the brains of aged APP/PS1 mice. (C) Quantification data show the percentage of TREM2+ area surrounding the 6E10+ A beta plaques (within 10μm from the border of the A beta plaques) in the brains of aged APP/PS1 mice with or without SCF+G-CSF treatment. (D) Representative orthographic view of z-stack images (12 z-stacks with 1μm intervals) displays the location and interaction of TREM2+/Iba1+ co-expressing cells (yellow) and 6E10+ A beta plaques (white) in the brains of APP/PS1 mice. (E) Quantification data show the percentage of TREM2+/Iba1+ co-expression area in the total of Iba1+ area in the vicinity of 6E10+ A beta plaques in the brains of aged APP/PS1 mice with or without SCF+G-CSF treatment. N=4-5. Mean ± SEM. * p<0.05 by Student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33269098), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse TREM2 by Immunocytochemistry/Immunofluorescence SCF+G-CSF treatment increases TREM2 expression in the Iba1+ microglia/macrophages surrounding the 6E10+ senile plaques. (A) Representative confocal images of TREM2 (red), 6E10 (purple) and Iba1 (green) triple immunofluorescence staining in the brains of aged APP/PS1 mice. Blue: Nuclear counterstaining by DAPI. (B) Representative orthographic view of z-stack images (12 z-stacks with 1μm intervals) illustrates the location and interaction of TREM2 + cells (red) and 6E10+ A beta plaques (white) in the brains of aged APP/PS1 mice. (C) Quantification data show the percentage of TREM2+ area surrounding the 6E10+ A beta plaques (within 10μm from the border of the A beta plaques) in the brains of aged APP/PS1 mice with or without SCF+G-CSF treatment. (D) Representative orthographic view of z-stack images (12 z-stacks with 1μm intervals) displays the location and interaction of TREM2+/Iba1+ co-expressing cells (yellow) and 6E10+ A beta plaques (white) in the brains of APP/PS1 mice. (E) Quantification data show the percentage of TREM2+/Iba1+ co-expression area in the total of Iba1+ area in the vicinity of 6E10+ A beta plaques in the brains of aged APP/PS1 mice with or without SCF+G-CSF treatment. N=4-5. Mean ± SEM. * p<0.05 by Student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33269098), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Mouse TREM2 Antibody by Western Blot A beta oligomers induce TREM2 proteolysis and sTREM2 release, which then binds A beta oligomers, but R47H sTREM2 binds less. A, western blot of cell lysate and unprocessed supernatant (sTREM2) of HEK293 cells coexpressing human DAP12 and full-length N-terminally-tagged wild-type (WT) human TREM2 (FL-TREM2) 16 h after adding A beta oligomers. This blot and those for A beta monomers and fibrils are reproduced in Figure S5 for comparison. B, quantification of sTREM2 release from transfected HEK293 cells expressing wild-type (green line) and R47H TREM2 (red line). C, quantification of sTREM2 release from wild-type TREM2 expressing HEK293 cells induced by doses of A beta oligomers (red line), monomers (green line), or fibrils (blue line). For both (B and C) error bars = SEM; ∗p < 0.05 ∗∗p < 0.01 ∗∗∗p < 0.001, n = 3 independent experiments; one-way ANOVA with Tukey's post-hoc multiple comparisons test. D, example field of single-molecule TIRF imaging of mixture of A beta oligomers (green) and wild-type TREM2 ectodomain (red), where colocalized spots appear yellow. Scale bar: 1 micron. Magnified image of three sections of field at right. E, proportion of monomeric or oligomeric A beta colocalized with wild-type sTREM2. F, proportion of A beta oligomers colocalized with wild-type or R47H TREM2 ectodomain. For (E and F), error bars = SEM; ∗∗∗∗p < 0.0001, n = 3 independent preparations, each analyzed in nine fields each; two-tailed t-test of significance. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33823153), licensed under a CC-BY license. Not internally tested by R&D Systems.