pe anti human cd86 bu63  (Proteintech)

Journal: Journal of the National Cancer Center

Article Title: Overexpression of ornithine decarboxylase 1 mediates the immune-deserted microenvironment and poor prognosis in diffuse large B-cell lymphoma

doi: 10.1016/j.jncc.2024.10.001

Figure Lengend Snippet: In vitro experiments demonstrate the impact of ODC1 on macrophage polarization. (A) Flow cytometry analysis of CD206 + M2 macrophages and CD86 + M1 macrophages after co-culture of M0 THP-1 cells with DLBCL cell lines overexpressing or knocking down ODC1. (B) ELISA measurement of cytokine levels in the supernatants after co-culture with different DLBCL cell lines. (C) RT-PCR analysis of M1/M2 marker gene expression in macrophages after co-culture with different DLBCL cell lines. (D) Plasmid-mediated overexpression and knockdown of ODC1 in THP-1 cell lines, with western blotting (upper panel) and RT-PCR (lower panel) used to detect ODC1 protein expression and mRNA levels. (E) Flow cytometry analysis of CD206 + M2 macrophages and CD86 + M1 macrophages in THP-1 cells with ODC1 expression discrepancy. (F) ELISA measurement of cytokine levels in the supernatants from THP-1 cells with ODC1 expression discrepancy. (G) RT-PCR analysis of M1/M2 marker gene expression in macrophages from different groups. The unpaired t -test was used to assess statistical differences between two groups. * P < 0.05; ⁎⁎ P < 0.01; ⁎⁎⁎ P < 0.001; ⁎⁎⁎⁎ P < 0.0001.

Article Snippet: Subsequently, AF488 conjugated anti-human CD68 antibody (clone # Y1/82A, Elabscience), APC conjugated anti-mouse CD206 antibody (clone # C068C2, Elabscience), or PE conjugated anti-human CD86 antibody (clone # BU63, Elabscience) were added, and the cells were incubated in the dark at room temperature for 30 min. After centrifugation, the cells were resuspended in 200 μL of PBS with 1 % BSA and analyzed using flow cytometry.

Techniques: In Vitro, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Marker, Gene Expression, Plasmid Preparation, Over Expression, Knockdown, Western Blot, Expressing

Journal: Journal of Translational Medicine

Article Title: SIRPB1 regulates inflammatory factor expression in the glioma microenvironment via SYK: functional and bioinformatics insights

doi: 10.1186/s12967-024-05149-z

Figure Lengend Snippet: A , B SIRPB1 expression in astrocytoma and glioblastoma cell clusters. C Correlation of SIRPB1 with immune cell infiltration. D Association between macrophage infiltration and SIRPB1; Wilcoxon and Spearman tests. E SIRPB1 and macrophage correlation in TCGA-GBM via TIMER2.0. F Kaplan–Meier curves for OS of TCGA-GBMLGG with ICR-high. G Co-localization of SIRPB1 with TMEM119, CD86, and CD163 in glioma. H SIRPB1 levels in glioma and monocyte lines from CCLE. I SIRPB1 expression in human and mouse glioma, microglia, and monocyte lines

Article Snippet: The induced THP-1 macrophages were collected, Fc receptors were blocked by human Fc Receptor Blocking Solution (Maokangbio), stained with FITC Anti-Mouse/Human CD11b Antibody (Elabscience, clone:M1/70), 7-AAD (Elabscience), APC Anti-Human CD206 Antibody (Elabscience, clone:15–2) and PE Anti-Human CD86 Antibody (Elabscience, clone:BU63), and detected and analyzed by Beckman cytoflex flow cytometry.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: SIRPB1 regulates inflammatory factor expression in the glioma microenvironment via SYK: functional and bioinformatics insights

doi: 10.1186/s12967-024-05149-z

Figure Lengend Snippet: SIRPB1 Knockout and Macrophage Polarization. A T7E1 assay results. WT wild-type, NC negative control, PC positive control. B SIRPB1 and FLAG-cas9 expression in THP-1 lines. C Sanger sequencing of SIRPB1 WT and SIRPB1 KO . D Protein expression post-M1/M2 treatments. E mRNA levels of M1/M2 markers (* P < 0.05, ** P < 0.01, *** P < 0.001, Dunnett’s test). F Flow cytometry of CD11b, CD86, CD206 in THP-1 lines

Article Snippet: The induced THP-1 macrophages were collected, Fc receptors were blocked by human Fc Receptor Blocking Solution (Maokangbio), stained with FITC Anti-Mouse/Human CD11b Antibody (Elabscience, clone:M1/70), 7-AAD (Elabscience), APC Anti-Human CD206 Antibody (Elabscience, clone:15–2) and PE Anti-Human CD86 Antibody (Elabscience, clone:BU63), and detected and analyzed by Beckman cytoflex flow cytometry.

Techniques: Knock-Out, Negative Control, Positive Control, Expressing, Sequencing, Flow Cytometry