cd11b monoclonal antibody  (Bio-Rad)

Journal: Journal of Neuroinflammation

Article Title: Early immune responses to systemic inflammation in the postnatal mouse brain initiated by migrating macrophages and leptomeningeal fibroblasts

doi: 10.1186/s12974-025-03559-4

Figure Lengend Snippet: RT-qPCR data showing fold changes in Saa3 , Cxcl9 , Irg1 , and Lcn2 transcript expression levels of brain cells in response to systemic inflammation. The transcript expression of Saa3 in brain CD11b(+) cells of mice with systemic inflammation shows very marked upregulation compared with saline control mice from 4 to 72 h after LPS administration ( A ). Furthermore, Saa3 expression in brain CD11b(−) cells of mice with systemic inflammation shows a marked increase from 4 to 72 h after LPS administration ( B ). Systemic inflammation-activated changes in Cxcl9 transcript expression in CD11b(−) cells are extremely upregulated compared with saline control mice at 4 and 12 h, and thereafter they decrease gradually, but they are still significantly elevated up to 72 h after LPS administration ( D ). The degree of Cxcl9 upregulation in CD11b(+) cells of mice with systemic inflammation is much lower than in CD11b(-) cells, but still significantly elevated up to 72 h after LPS administration ( C ). The transcript expression of Irg1 in CD11b(+) cells of mice with systemic inflammation shows the greatest increase compared with saline control mice 4 h and 12 h after LPS administration. Thereafter, expression decreases gradually, but it remains significantly elevated up to 48 h after LPS administration ( E ). Irg1 expression in CD11b(−) cells of mice with systemic inflammation also shows marked upregulation 4 and 12 h after LPS treatment, followed by a gradual decrease ( F ). Systemic inflammation-activated changes in Lcn2 transcript expression levels in CD11b(−) cells are highly upregulated compared with saline control from 4 to 24 h after LPS administration and decrease gradually thereafter ( H ). The degree of Lcn2 upregulation is much lower in CD11b(+) cells than in CD11b(-) cells ( G ). Mean ± SEM. p < 0.05 and ** p < 0.01 (see Methods-Statistical analysis), compared with saline control

Article Snippet: The pellets were resuspended in cold D-PBS (+) containing 0.5% BSA and incubated with R-phycoerythrin (PE)-conjugated primary human/mouse CD11b monoclonal antibody (130–113–235, Miltenyi Biotec) and Fc receptor blocking reagent (130–092–575, Miltenyi Biotec), followed by incubation with MicroBeads UltraPure conjugated to anti-PE monoclonal antibody (130–105–639, Miltenyi Biotec).

Techniques: Quantitative RT-PCR, Expressing, Saline, Control

Journal: bioRxiv

Article Title: Inflammation-enhanced synapse-specific phagocytosis by adult APP microglia in a microfluidic neuron–microglia co-culture model

doi: 10.1101/2025.08.19.671013

Figure Lengend Snippet: (A) Representative mosaic image from the live PSD95 uptake assay (left). Images show WT and APP microglia under vehicle or LPS (100 ng/mL). TdTomato + microglia (red) interact with PSD95-EGFP + neuronal structures (green). White outlines indicate microglial ROIs used for quantification. (B) Quantification of PSD95 internalization per microglial cell. LPS selectively enhanced PSD95 uptake in APP microglia, suggesting increased synaptic targeting. One-way ANOVA with Tukey’s post hoc test. (C) CD11b blockade reduced PSD95 internalization in LPS-treated APP microglia, supporting a partial dependence on CD11b signaling. (D) Uptake of pHrodo™ Green Zymosan, a non-specific substrate, remained unchanged, indicating that the phagocytic increase was specific to synaptic material. (E) Representative immunostaining of synaptic compartments shows Synaptophysin (SYP, green) and Homer1 (red) puncta. (F) Quantification of synaptic connectivity, defined as the proportion of SYP puncta assigned by Homer1. No differences were observed between conditions, suggesting that LPS did not disrupt overall synaptic architecture. To avoid local effects of microglial contact, analysis was restricted to regions ≥75 μm from the nearest microglial soma. Data in (B–D, F) represent mean ± SEM, normalized to vehicle of each genotype; n = 4–6 coverslips per condition from ≥3 independent experiments. Scale bars: (A) 500 µm (left), 100 µm (right); (E) 20 µm.

Article Snippet: To block CD11b, the anti-CD11b monoclonal antibody (clone M1/70.15; 10 μg/mL; Bio-Rad, MCA74EL) was added 24 h prior to LPS treatment and maintained throughout the stimulation and recovery phases.

Techniques: Immunostaining

Journal: iScience

Article Title: Dietary fermentable polyols fuel gut inflammation through M1 macrophage polarization and gut microbiota

doi: 10.1016/j.isci.2025.112934

Figure Lengend Snippet: Sorbitol intake promotes M1 macrophage accumulation in the colonic lamina propria and exacerbates colitis in an IL-1β-dependent manner (A) The proportion of M1 or M2 macrophage (Ly6C + /CD206 + , CD45 + , FVS780 - , Siglec-F - , Ly6G − , CD11b + , F4/80 + ) in colonic lamina propria (cLP) ( n = 8 or 9 samples per group). (B) The representative flow cytometry plots of M1 or M2 macrophage ( n = 8 or 9 samples per group). (C) Right: the survival rate in WT, IL-1β KO mice with or without sorbitol intake after DSS treatment ( n = 7 samples per group). Left: the survival days post-DSS treatment ( n = 7 samples per group). Plots represent the mean ± SEM. Welch’s t test (B), log rank test (C), and Sidak’s multiple comparisons tests (C). ∗∗ p < 0.01 and ∗ p < 0.05.

Article Snippet: PE, CD11b Monoclonal Antibody (M1/70) , Thermo Fisher Scientific , Cat# 12-0112-83; RRID: AB_273487005.

Techniques: Flow Cytometry

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir treatment ameliorates liver damage and fibrosis in MCD diet-induced MASH mouse model. A: Schematic illustration of the experimental workflow for the daclatasvir treatment strategy in a methionine-choline deficient (MCD) diet-induced MASH mouse model. B: Liver weight, LW/BW, and LW/BMI were recorded for MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. C: Representative images of H&E, PSR, and CD11b staining of liver samples from vehicle or daclatasvir treated mice at 4 weeks of MCD diet administration. Scale bar, 200 μm for H&E and PSR; 100 μm for CD11b. D: Quantitative results for PSR and CD11b staining shown in (C). E: Enzyme activities of serum hepatic transaminases in MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. F–H: The mRNA expression of genes associated with lipid catabolism (F), inflammation (G), and fibrosis (H) in liver tissues from MCD diet-induced MASH mice following 3 weeks of daclatasvir or vehicle treatment. For (B–H), n = 6 mice per group. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; Student’s t test for statistics.

Article Snippet: The sections were incubated with a rabbit anti-CD11b primary antibody (BM3925, Boster) overnight at 4°C, followed by incubation with a goat anti-rabbit fluorophore-conjugated secondary antibody (A-11036, Invitrogen).

Techniques: Staining, Expressing