anti human cd88  (Bio-Rad)

Journal: STAR Protocols

Article Title: Protocol for isolating cervico-vaginal fluid cells from Macaca mulatta to study immunological and functional changes during pregnancy

doi: 10.1016/j.xpro.2025.103927

Figure Lengend Snippet: Gating strategy for flow cytometry phenotype analysis of CVL cells (A) Filter the CVL cell solution in a FACS 5 ml polystyrene round-bottom tube with cell-strainer cap. (B) After centrifugation, the cells pellet is visible. (C) After incubating the cells with the Ab mix, add 500 μl/tube of cold FACS washing buffer. (D) Representative density plots of Rhesus CVL samples. Forward scatter area (FSC-A) and forward scatter height (FSC-H) gating on cells to exclude doublets. Dead cells were excluded by Live/Dead Fixable Aqua Dead Cell Stain kit. FSC and side (SSC) scatter gating on cells to exclude cellular debris. CD45 − cells gate representing non-leukocyte cells and CD45 + cells positive gating representative leukocyte cells. Inside the CD45 + cells, the leukocyte subpopulations were gated as monocytes/macrophages (CD3 − HLA-DR + CD19 − CD20 − CD14 + ); activated B cells (CD3 − HLA-DR + CD14 − CD19 + CD20 + ); neutrophils (CD3 − CD14 low HLA-DR − CD88 + CD56 − ); NK cells (CD3 − CD14 − HLA-DR − CD56 + ); resting B cells (CD3 − HLA-DR − CD14 − CD56 − CD19 + CD20 + ); T cells (CD14 − CD56 − CD3 + ); and NKT cells (CD14 − CD3 + CD56 + ); myeloid dendritic cells (mDCs, CD3 − CD14 − HLA-DR − CD56 − CD19 − CD20 − CD123 − ); plasmacytoid dendritic cells (pDCs, CD3 − CD14 − HLA-DR − CD56 − CD19 − CD20 − CD123 + ). A similar gating strategy can be used to characterize human cells, with the only difference that CD66b should be used instead of CD88 for neutrophils. (E) Example of data analysis: frequency of neutrophils in CVL at different time points during gestation (before E. coli injection = gestational age 140 days; delivery = gestational age gestational age 146 days).

Article Snippet: Anti-human CD88 (clone MCA2059A647T) – dilution 1:10 , Bio-Rad , Cat# MCA2059A647T, RRID: AB_1102420.

Techniques: Flow Cytometry, Centrifugation, Staining, Injection

Journal: Journal of histotechnology

Article Title: C5aR expression in kidney tubules, macrophages and fibrosis.

doi: 10.1080/01478885.2024.2430041

Figure Lengend Snippet: Figure 1. C5aR was detected in representative sections from non-diseased human kidney by IHC with 3 different C5aR monoclonal antibodies: 8D6 (a), S5/1 (b and d), and P12/1 (c). Red dotted circles and arrows highlight examples of tubules and interstitial (or glomerular) cells, respectively, that are detected by the indicated antibody (brown). (e) Section incubated with mouse IgG2a antibody as an isotype control for S5/1 and P12/1. (f) Section incubated with rat IgG2b antibody as an isotype control for 8D6. (a), (b), and (f) are adjacent serial sections, as are (c), (d), and (e). Images were counterstained for cell nuclei and the scale bar in (a) corresponds to 100 µm for all images.

Article Snippet: For IHC, which was performed manually, the following C5aR monoclonal anti-human antibodies were used: clone S5/1, mouse IgG2a (Hycult Biotech [Wayne PA, U.S.A.] Cat# HM2094, RRID: AB_533292), clone P12/1, mouse IgG2a (Bio-Rad [Hercules, CA, U.S.A.] Cat# MCA2059, RRID: AB_566906), clone 8D6, rat IgG2a (Thermo Fisher Scientific [Waltham, MA, U.S.A.] Cat# MA1-70060, RRID:AB_1073841).

Techniques: Bioprocessing, Incubation, Control