horseradish peroxidase conjugated goat anti rat igg  (Novus Biologicals)

Journal: Bioactive Materials

Article Title: Countering postoperative immune suppression with a self-assembling dendritic cell nanovaccine

doi: 10.1016/j.bioactmat.2026.05.005

Figure Lengend Snippet: BAITs promote DC activation and antigen presentation via enhanced lysosomal processing. (A) Schematic for MHC Ⅰ/Ⅱ immunostaining. DCs were treated with antigens or BAITs (12 h), then incubated in fresh medium (with TGFβ) for 24 h before staining. (B, C) Quantification of surface MHC Ⅰ (B) and MHC Ⅱ (C) on DCs (n = 8; n.s. is P > 0.05, ∗∗∗ is P < 0.001, ∗∗∗∗ is P < 0.0001 by one-way ANOVA with Bonferroni post-hoc test). (D, E) Representative immunofluorescence images of DCs (blue: nuclei; yellow: LAMP1; green: MHC Ⅰ; magenta: MHC Ⅱ). BAIT-treated DCs show strong surface MHC and minimal intracellular antigen signal, whereas antigen-treated DCs show retained antigen (red) and weak surface MHC signal. (F–H) Flow cytometric analysis of MHC Ⅰ (F), MHC Ⅱ (G), and CD80 (H) expression on murine CD11c + DCs after exposure to antigens or BAITs. (I) Quantification of MHC Ⅰ, MHC Ⅱ, and CD80 expression in DCs by flow cytometry (n = 3; ∗∗∗ is P < 0.001, ∗∗∗∗ is P < 0.0001 by two-way ANOVA with Bonferroni post-hoc test). (J) Heatmap of DC activation-related gene expression after treatment with antigens or BAITs, highlighting upregulation of maturation and antigen presentation genes and downregulation of immunosuppressive genes by BAITs (n = 3). Data are presented as mean ± SD.

Article Snippet: After treatment, the cells were washed twice with PBS and fixed in 4% paraformaldehyde for 20 min. For intracellular protein staining, fixed cells were permeabilized with 0.1% Triton X-100 at room temperature for 10 min. Next, the cells were blocked with 2% BSA and incubated with rat CoraLite Plus 488 anti-mouse LAMP1 antibody (1:200) and rabbit anti-mouse pSAP (1:200) antibody overnight at 4 °C.

Techniques: Activation Assay, Immunopeptidomics, Immunostaining, Incubation, Staining, Immunofluorescence, Expressing, Flow Cytometry, Gene Expression

Journal: Bioactive Materials

Article Title: Injection site dictates the immune response to a biodegradable polymer and corresponding collagen regeneration

doi: 10.1016/j.bioactmat.2026.04.004

Figure Lengend Snippet: ID tissue exhibits higher densities of fibroblasts, macrophages, and microvessels compared to SC tissue. Immunofluorescence analysis comparing the expression of Vimentin (a fibroblast marker), CD68 (a macrophage marker), and CD31 (an endothelial cell marker) in ID and SC tissues. For each group, n = 3; data represent mean ± s.d.; ∗P < 0.05 and ∗∗P < 0.01.

Article Snippet: After antigen retrieval and blocking, sections were incubated overnight at 4 °C with primary antibodies targeting vimentin, CD68 (ABclonal, A20803), CD31 (R&D SYSTEMS, AF3628), HMGB1 (Cell Signaling Technology, 3935), HSP70 (ABclonal, A23457), NF-κB p65 (ABclonal, A19653), CD206 (Cell Signaling Technology, 24595), and FAPα (ABclonal, A23789 ).

Techniques: Immunofluorescence, Expressing, Marker

Journal: STAR Protocols

Article Title: Protocol for isolating and culturing microglia from the adult mouse brain using a magnetic-activated cell sorting system

doi: 10.1016/j.xpro.2026.104471

Figure Lengend Snippet: Representative workflow of isolation and culture of adult mouse microglia (A and B) Dissect the brain into small pieces on ice in Petri dish. (C) Collect cell pellets in C-tubes following mechanical/enzymatic dissociation using a gentleMACS dissociator. (D) Preparation of cell straining and debris removal processes. (E) Perform debris removal by carefully overlaying ice-cold PBS onto the cell suspension. (F) After centrifugation, identify three layers; the middle, yellowish layer corresponds to myelin dna debris. (G) Enrich CD11b+ cells by magnetic separation using appropriate columns. (H) Seed isolated microglia onto 6-wll culture plates for downstream assays.

Article Snippet: CD11b Microbead (Clone: M1/70.15.11.5) (1:10 ratio) , Miltenyi Biotech , Cat#130-049-601; RRID: AB_2927911.

Techniques: Isolation, Suspension, Centrifugation

Journal: STAR Protocols

Article Title: Protocol for isolating and culturing microglia from the adult mouse brain using a magnetic-activated cell sorting system

doi: 10.1016/j.xpro.2026.104471

Figure Lengend Snippet: Flow cytometry for MACS-isolated microglia purity (A and B) Total CD11b-positive cells from MACS columns; (C) Cell viability assessed by Zombie Red staining; (D) Infiltrating leukocytes, including neutrophils (Ly6G+) and T lymphocytes (CD3+); (E) Proportions of microglia (CD11b+CD45low) versus monocytes/border-associated macrophages (CD11b+CD45high); (F) Proportion of resting (homeostatic) microglia identified as CD11b+TMEM119+ cells.

Article Snippet: CD11b Microbead (Clone: M1/70.15.11.5) (1:10 ratio) , Miltenyi Biotech , Cat#130-049-601; RRID: AB_2927911.

Techniques: Flow Cytometry, Isolation, Staining