We used BD Stemflow Human MSC Analysis Kit where the following antibodies were provided, CD73-APC, CD105-PerCP-Cy5.5, CD90-FITC, CD44-PE and the PE channel open to use in combination with the supplied negative MSC cocktail (PE CD45, PE CD34, PE CD11b
, PE CD19 and PE HLA-DR) and also included antibody conjugate.
In this study, investigators looked at the integrin CD11b
, which I present on myeloid cells and normally helps myeloid cell migration and its ability to fight disease.
Lineage-specific markers, including myeloid (CD13, CD33, CD117, MPO), monocytic (CD11b
, CD14, CD64), megakaryocytic (CD61), B-cell (CD19, CD10, CD20) and T-cell (CD3) antigens, were negative.
The gating strategy for analyzing interstitial macrophages was the same as in previous reports with some modifications.,, In brief, the isolated cells were incubated with PE/Cy5-F4/80, PE-CD206, PE/Cy7-CD11c antibodies, and CD11b
antibodies (BioLegend, San Diego, CA, USA) at the concentrations recommended in the manufacturer's protocol [Supplementary Figure 1 [SUPPORTING:1]].
CD33, CD14, CD11b
, HLA DR, and myeloperoxidase were positive and CD34 was negative in flow cytometric analysis.
The differentiation of THP-1 cells into macrophage-like cells was determined by using FITC anti-human CD11b
antibody (BioLegend, 301329), which stains macrophages, and FITC anti-human CD14 antibody (BioLegend, 301803), which stains monocytes.
With suspicion of LAD it was decided to investigate it and diagnosis was confirmed by flow cytometry showing absence of CD18, CD11b
, CD11c on neutrophils.
Pienta, "Prostate cancer promotes CD11b
positive cells to differentiate into osteoclasts," Journal of Cellular Biochemistry, vol.
For this, we assessed expression of surface markers TLR4, CD11b
, and CD163 on circulating monocytes as well as production of TNF[alpha] and IL-10 by whole blood cells in the presence and absence of LPS.
After intravitreal injection with 1 f g/f l AC220 (or control vehicle), mouse retinas at P15, P18, and P21 (6-10 retinas per group) were obtained for the isolation of retinal cells as described previously, using antimouse CD11b
magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany) and a premoistened MS column (BD Biosciences) .
Antibodies against SP, GFAP, and CD11b
(OX42) and rabbit anti-mouse secondary antibody were also purchased from Abcam, England.