goat anti human trem2 polyclonal microglia  (R&D Systems)

Journal: bioRxiv

Article Title: Increased CD33 levels tune activation and function of induced human microglial cells through inhibition of the TREM2 pathway

doi: 10.64898/2026.01.28.701050

Figure Lengend Snippet: (A) Schematic of siRNA transfection of iMG. Cells were transfected 24 hours after iMG plating. After an additional 24 hours, cells were incubated with 2 μg/mL of pHrodo-labeled oligomerized amyloid (1-42) and imaged over a 20-hour window. (B) Confocal fluorescence images of iMG assessed for internalization of pHrodo-oligomerized amyloid beta treated as in (A). Digital phase contrast was used to visualize iMG. Images are stitched from four 20X images. Scale bar represents 400 μm. (C) Normalized intensity of internalized pHrodo-oligomerized amyloid beta in iMG per hour. Data are normalized to mock transfected control. (D) Normalized intensity of internalized pHrodo-oligomerized amyloid beta in iMG at endpoint. Data are normalized to mock transfected control. (E) RNA isolated from iMG 24 hours after transfection was used to quantify the percent CD33 mRNA expression by qPCR. (F) Change in TREM2 secretion in conditioned media from iMG was quantified 24 hours after transfection by MSD. (G) Protein lysates isolated from iMG 24 hours after transfection were used to quantify percent SYK phosphorylation by AlphaLISA. Data represent mean ± SEM using one-way ANOVA with uncorrected Fisher’s LSD (D) and Dunnett’s multiple comparison test (E-G). Data points (n-numbers) are plotted on each bar graph and scatter plot each representing an independent experimental replicate. Line plots are representative of three independent experimental replicates (C).

Article Snippet: Briefly, streptavidin-coated small spot MSD plates (Meso Scale Diagnostics, Rockville, MD) were coated with Biotinylated goat anti-human TREM2 polyclonal antibody (R&D Systems) used as a capture antibody for 1.5 hours with agitation at 600 rpm.

Techniques: Transfection, Incubation, Labeling, Fluorescence, Control, Isolation, Expressing, Phospho-proteomics, Comparison

Journal: bioRxiv

Article Title: Increased CD33 levels tune activation and function of induced human microglial cells through inhibition of the TREM2 pathway

doi: 10.64898/2026.01.28.701050

Figure Lengend Snippet: (A) Schematic of AAV6-mediated transduction of iMG. Cells were transduced with an MOI of 125000 at the time of plating. On day 3, cells were incubated with 2 μg/mL of pHrodo-labeled oligomerized amyloid (1-42) and imaged over a 20-hour window. (B) Protein lysates isolated from iMG 72 hours after transduction were used to quantify percent CD33 protein expression by MSD. Data are normalized to no AAV6 transduction control. (C) Confocal fluorescence images of iMG assessed for internalization of pHrodo-oligomerized amyloid beta treated as in (A). Digital phase contrast was used to visualize iMG. Images are stitched from four 20X images. Scale bar represents 400 μm. (D) Normalized intensity of internalized pHrodo-oligomerized amyloid beta in iMG transduced with or without AAV6-CD33 or AAV6-mCherry per hour. Data are normalized to no AAV6 transduction control. (E) Normalized intensity of internalized pHrodo-oligomerized amyloid beta in iMG transduced with or without AAV6-CD33 or AAV6-mCherry at endpoint. Data are normalized to no AAV6 transduction control. (F) TREM2 secretion in conditioned media was quantified from iMG 72 hours after transduction by MSD. (G)(left) Percent change in TREM2 secretion in conditioned media was quantified from HMC3 cells 48 hours after transduction with increasing AAV6-CD33 concentrations by MSD. Data are represented as a percent change relative to dose-matched AAV6-mCherry. (right) TREM2 secretion in conditioned media was quantified from HMC3 cells 48 hours after transduction of AAV6-CD33 by MSD. (MOI = 250000). (H) Protein lysates isolated from HMC3 cells 48 hours after transduction were used to quantify percent full length TREM2 protein expression by MSD. (MOI = 250000). Data represent mean ± SEM using one-way ANOVA with post-hoc Tukey’s multiple comparisons test (B), with uncorrected Fisher’s LSD (E), with post-hoc Dunnett multiple comparisons test (F, G) and one-tailed unpaired t-test (H). Data points (n-numbers) are plotted on each bar graph and scatter plot each representing an independent experimental replicate. Line plots represent three independent experimental replicates (D, G).

Article Snippet: Briefly, streptavidin-coated small spot MSD plates (Meso Scale Diagnostics, Rockville, MD) were coated with Biotinylated goat anti-human TREM2 polyclonal antibody (R&D Systems) used as a capture antibody for 1.5 hours with agitation at 600 rpm.

Techniques: Transduction, Incubation, Labeling, Isolation, Expressing, Control, Fluorescence, One-tailed Test

Journal: bioRxiv

Article Title: Increased CD33 levels tune activation and function of induced human microglial cells through inhibition of the TREM2 pathway

doi: 10.64898/2026.01.28.701050

Figure Lengend Snippet: (A) Schematic of the TREM2-CD33 pathway. CD33 inhibition acts on SYK activation, upstream of microglial activation pathways AKT, ERK, p38, and JNK. (B) Protein lysates isolated from THP-1 monocytes 48 hours after transduction were used to quantify percent SYK phosphorylation by MSD. (MOI = 31250) (C) 48 hours post-transduction, THP-1 cells (MOI = 31250) were treated with increasing concentrations of TREM2-targeting antibody agonist (0-10 μg/mL). After 5 minutes, protein lysates were isolated and used to quantify percent SYK phosphorylation by MSD. (D) 48-hours post-transduction of increasing concentrations of AAV6-CD33 (MOI = 0-31250), THP-1 cells were treated with a TREM2-targeting antibody agonist (2.5 ug/mL). After 5 minutes, protein lysates were isolated and used to quantify percent SYK phosphorylation by MSD. (E) Percent AKT and GSK3β phosphorylation was quantified from HMC3 cell lysates 48 hours after transduction of AAV6-CD33, quantified by MSD and ELISA, respectively. (F) Percent ERK and p38-MAPK phosphorylation was quantified from HMC3 cell lysates 48 hours after transduction of AAV6-CD33, quantified by ELISA. (G) Percent JNK phosphorylation was quantified from HMC3 cell lysates 48 hours after transduction of AAV6-CD33, quantified by ELISA. (H) (left to right) Percent expression of secreted IL10, IL1β, TNFα, and IL6 in conditioned media from HMC3 cells 24 hours after LPS challenge and 48 hours after transduction of AAV6-CD33 was quantified by MSD. Data represent mean ± SEM using one-tailed unpaired t-test (A, E-G), and two-way ANOVA using uncorrected Fisher’s LSD with a single pooled variance (H). Data points (n-numbers) are plotted on each bar graph and scatter plot each representing an independent experimental replicate. Line plots are representative of three and two independent experimental replicates (C, D, respectively).

Article Snippet: Briefly, streptavidin-coated small spot MSD plates (Meso Scale Diagnostics, Rockville, MD) were coated with Biotinylated goat anti-human TREM2 polyclonal antibody (R&D Systems) used as a capture antibody for 1.5 hours with agitation at 600 rpm.

Techniques: Inhibition, Activation Assay, Isolation, Transduction, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Expressing, One-tailed Test

Journal: mAbs

Article Title: Building a potent TREM2 agonistic, biparatopic, common light chain antibody

doi: 10.1080/19420862.2025.2546554

Figure Lengend Snippet: TREM2 tool antibodies activity in various formats. (a) BMDM viability assay with TREM2 mAbs and isotype control in solution (5 day incubation, 0.2 nM CSF-1 with 100 nM antibodies or media only), (b) same mAbs platebound in BMDM viability assay. Red dashed line indicates viability of cells with isotype control in both assays. (c) Alternate antibody formats evaluated include asymmetric bispecific (AsBs), tetravalent (Tv), symmetric tetravalent bispecific (SymTvbs) with Efab domain substitution indicated in light gray, and asymmetric tetravalent bispecific (AsTvBs). Bispecifics were generated by combining antibodies 3 or 4 with antibody 10. (d) BMDM viability assay of alternate formats compared to soluble mAbs (48 h incubation, no CSF-1 with antibodies at 100 nM concentration or media only). Red dashed line indicates viability of cells with isotype control. (e) Schematic of NanoBiT huTrem2/hudap12/husyk assay developed to enable high-throughput screening of antibodies for their ability to trigger DAP12 phosphorylation, resulting in an interaction with SYK, in a human system. (f) NanoBiT pDAP12 assay validation with soluble and crosslinked antibodies at 50 nM, with and without huTREM2 expression. (g) Dose-response curves of mAbs and alternate formats in NanoBiT pDAP12 assay, normalized to Emax of plate control AsTvBs 4/10 @ 200 nM. (h) Schematic of hypothetical binding mode of a tetravalent bispecific with TREM2 shown in cyan and gray, TREM2 biparatopic antibody shown in black and blue, and cell membrane shown in pink.

Article Snippet: A 96-well small spot streptavidin gold MSD plate was incubated with a biotinylated polyclonal goat anti-TREM2 antibody (R&D Systems #BAF1828) overnight at 4°C with shaking at 500 RPM.

Techniques: Activity Assay, Viability Assay, Control, Incubation, Generated, Concentration Assay, High Throughput Screening Assay, Phospho-proteomics, Biomarker Discovery, Expressing, Binding Assay, Membrane

Journal: mAbs

Article Title: Building a potent TREM2 agonistic, biparatopic, common light chain antibody

doi: 10.1080/19420862.2025.2546554

Figure Lengend Snippet: (a) Common light chain (cLC) antibody campaign overview. Red dots indicate number of CDRs involved in affinity maturation at each stage. (b) Model of membrane-bound TREM2 ECD including IgV (resi 19–134, green) from PDB 5ELI and stalk (resi 135–174, orange line) with sheddase site between His157 and Ser158 indicated by dashed line. (c) Family tree of select antibodies from cLC campaign. Generation 1 = antibodies from screen; subsequent generations: 2 = H1/H2 shuffle; 3 = H3 mutagenesis, 4: H1/H2/H3 mutagenesis using walking oligos. Monovalent binding affinity (K D ) reported for antibodies with measurable value as determined by surface plasmon resonance analysis of antibodies using single-cycle analysis and kinetic parameters determined using steady state or 1:1 kinetics model.

Article Snippet: A 96-well small spot streptavidin gold MSD plate was incubated with a biotinylated polyclonal goat anti-TREM2 antibody (R&D Systems #BAF1828) overnight at 4°C with shaking at 500 RPM.

Techniques: Membrane, Mutagenesis, Binding Assay, SPR Assay

Journal: mAbs

Article Title: Building a potent TREM2 agonistic, biparatopic, common light chain antibody

doi: 10.1080/19420862.2025.2546554

Figure Lengend Snippet: Characterization of the lead antibodies combined in SymTvBs format. (a) SymTvBs reformatting of lead pair 7409/7268 into two antibody orientations. (b, c) Dose-response curves in NanoBiT and THP1 assays, 200 nM nM AsTvBs 4/10 used for normalization. (d) Viability in hTREM2 BMDMs, derived from human TREM2 bac transgenic mice, with antibodies tested at 20 and 1 nM, normalized to 200 nM AsTvBs. (e, f) Activity in THP1 pSYK AlphaLISA and aggregation assay, normalized to 31.6 nM undefined 4/10. (g) THP1 pSYK specific for pTyr525/526 by AlphaLISA with 20 nM antibody with or without 0.5 mg/mL liposomes (* p < 0.05; ** p < 0.01).

Article Snippet: A 96-well small spot streptavidin gold MSD plate was incubated with a biotinylated polyclonal goat anti-TREM2 antibody (R&D Systems #BAF1828) overnight at 4°C with shaking at 500 RPM.

Techniques: Derivative Assay, Transgenic Assay, Activity Assay, Liposomes

Journal: mAbs

Article Title: Building a potent TREM2 agonistic, biparatopic, common light chain antibody

doi: 10.1080/19420862.2025.2546554

Figure Lengend Snippet: Crystal structures of Fabs 7268 and 7411 in complex with TREM2. (a) Side view of the Fab 7268 (heavy chain, magenta; light chain, turquoise) bound to MBP-TREM2 IgV (gray and green). (b) Close-up view of the Fab 7268/TREM2 IgV binding interface, highlighting key residues within 4 Å of each other. (c) Side view of the Fab 7411 (heavy chain, purple; light chain, marine) in complex with the TREM2 stalk peptide (residues 131–148, green). (d) Detailed view of the Fab 7411/TREM2 stalk peptide interface, illustrating the TREM2 stalk wrapping around the CDR H3 residues of Fab 7411.

Article Snippet: A 96-well small spot streptavidin gold MSD plate was incubated with a biotinylated polyclonal goat anti-TREM2 antibody (R&D Systems #BAF1828) overnight at 4°C with shaking at 500 RPM.

Techniques: Binding Assay