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: Science advances

Article Title: Senolytic treatment for low back pain.

doi: 10.1126/sciadv.adr1719

Figure Lengend Snippet: Fig. 5. Senolytics reduced SnCs and pain-related neuroplastic changes in the spinal cord of sparc−/−mice. (A) Spinal cords from perfused animals treated with seno- lytic drugs or vehicle were collected and processed for histological evaluation. Representative images showing higher p16Ink4a immunoreactivity in the dorsal horn (delin- eated in dashed line) in sparc−/− compared to that in wild-type mice at 9 months. (B) Quantification of the increased immunoreactivity (percentage area) (% Area IR). (C) p16Ink4a immunoreactivity was reduced in treated sparc−/− animals. The spinal cords were also assessed for pain-related neuroplastic changes. (D) Representative images of calcitonin gene-related peptide (CGRP), glial fibrillary acidic protein (GFAP), and CD11b immunoreactivity in the dorsal horn of sparc−/− mice (delineated in a dashed line). The immunoreactivity (percentage area) was quantified for (E) CGRP, (F) GFAP, and (G) CD11b. The % area with immunoreactivity above a set threshold was calcu- lated from the total area of the dorsal horn. n = 7 to 8 animals per group (3 to 4 males and 4 females). DAPI was used as a counterstain. The average of three separate images was calculated for each animal and used to calculate the mean of the treatment group. Scale bars (A and D), 100 μm. Data are presented as means ± SD and were analyzed by a two-tailed t test or an ordinary one-way ANOVA followed by Tukey’s post hoc test. */#P < 0.05, **P < 0.01, ***/###P < 0.001, and ****P < 0.0001. * indicates a significant difference compared with sparc−/−, and # indicates a significant difference compared with single-drug treatment.

Article Snippet: Slides were then incubated with either recombinant anti- CDKN2A/p16Ink4a antibody (1:100; Abcam, catalog no. ab211542), sheep anti- CGRP polyclonal antibody (1:1000; Enzo Life Sciences, catalog no. BMLCA11370100, lot no. 0807B74), goat anti- GFAP polyclonal antibody (1:1000; Sigma- Aldrich, catalog no. SAB2500462, lot no. 747852C 2G2), or rat monoclonal anti- CD11b antibody (1:1000; Bio- Rad, catalog no. MCA711G, lot no. 0614) in blocking buffer overnight at 4°C; washed three times for 5 min in PBS; and incubated for 1.5 hours at room temperature with appropriate donkey- derived secondary antibodies from Jackson ImmunoResearch: donkey anti- sheep Cy3, catalog no. 713- 165- 147; donkey anti- goat Alexa Flour 594, catalog no. 705- 85- 144; donkey anti- rat Alexa Fluor 488, catalog no. 712- 225- 153; donkey anti- rabbit Alexa Fluor 488, catalog no. 711- 545- 152; and donkey anti- rabbit Cy3, catalog no. 711- 165- 152 in blocking buffer.

Techniques: Two Tailed Test