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dc marker  (Miltenyi Biotec)


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    Miltenyi Biotec dc marker
    Dc Marker, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd209/CD209+(DC-SIGN)+Antibody%2C+anti-human%2C+REAfinity/pmc12681927-309-9-18
    Average 93 stars, based on 17 article reviews
    dc marker - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Fluorescence:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20™ Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Method Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), 20, CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) Any of human-derived CD14 positive cell lines (THP-1, U937, and human-derived iPS-CD14 positive cell were used here) and NKT cell ligand RK-163 were cultured for 8-48 hr in the presence or absence of GM-CSF to prepare “RK-163-pulsed human CD14 positive cell line”. ..

    FACS:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20™ Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Method Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), 20, CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) Any of human-derived CD14 positive cell lines (THP-1, U937, and human-derived iPS-CD14 positive cell were used here) and NKT cell ligand RK-163 were cultured for 8-48 hr in the presence or absence of GM-CSF to prepare “RK-163-pulsed human CD14 positive cell line”. ..

    Centrifugation:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20™ Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Method Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), 20, CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) Any of human-derived CD14 positive cell lines (THP-1, U937, and human-derived iPS-CD14 positive cell were used here) and NKT cell ligand RK-163 were cultured for 8-48 hr in the presence or absence of GM-CSF to prepare “RK-163-pulsed human CD14 positive cell line”. ..

    Expressing:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20™ Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Method Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), 20, CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) Any of human-derived CD14 positive cell lines (THP-1, U937, and human-derived iPS-CD14 positive cell were used here) and NKT cell ligand RK-163 were cultured for 8-48 hr in the presence or absence of GM-CSF to prepare “RK-163-pulsed human CD14 positive cell line”. ..

    Flow Cytometry:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20™ Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Method Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), 20, CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) Any of human-derived CD14 positive cell lines (THP-1, U937, and human-derived iPS-CD14 positive cell were used here) and NKT cell ligand RK-163 were cultured for 8-48 hr in the presence or absence of GM-CSF to prepare “RK-163-pulsed human CD14 positive cell line”. ..

    Concentration Assay:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Cell Culture:

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20TM Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) For preparation of the cells obtained by culturing human CD14 positive cells (monocytes) immediately after separation using anti-CD14 antibody and human CD14 negative cells or human Elu-CD14 positive cells (monocytes) for 2 days with GM-CSF and NKT cell ligand RK-163 (respectively called “RK-163-pulsed human CD14 positive cell”, “RK-163-pulsed human CD14 negative cell”, “RK-163-pulsed human Elu-CD14 positive cell”), and the dendritic cells induced to differentiate in 2)-(2) by culturing for 6 days with addition of GM-CSF and IL-4 (called “RK-163-pulsed human dendritic cell”), RK-163 (bulk powder was dissolved in PBS added with DMSO and 0.5% Tween 20) was added to each cell to a final concentration of 100 ng/mL, and the cells were cultured at 37° C., 5% CO2 for 48 hr to produce “RK-163-pulsed cells”. ..

    Article Title: Technology for efficient activation of NKT cells
    Article Snippet: 3) Analysis Method (1) Calculation Method of Viable Cell Number and Survival Rate After staining with 0.4% Trypan Blue (Bio-Rad), unstained viable cells and blue-stained dead cells were measured twice with TC20™ Automated Cell Counter (Bio-Rad). .. The average thereof was calculated and the viable cell number and survival rate were calculated. (2) Fluorescence Activated Cell Sorting (FACS) Analysis Method Phycoerythrin-labeled anti-human antibody against each of the following antigens [CD14 (clone HCD14, Biolegend), CD11c (clone 3.9, Biolegend), CD11b (clone ICRF44, BD Biosciences), CD45 (clone HI30, Biolegend), HLA-DR (clone L243, Biolegend), CD1d (clone 51.1, Biolegend), CD40 (clone 5C3, Biolegend), CD95(Fas) (clone DX2, Biolegend), 20, CD80 (clone 2D10, Biolegend), CD86 (clone IT2.2, Biolegend), and CD209 (clone DC-SIGN, Miltenyi Biotech)] was added to the cells, and static reaction was performed under shading for 30 min at 4° C. After centrifugation and washing, the cells were suspended in 0.5% human serum albumin-added PBS, and the purity of CD14 positive cells and expression of various surface antigens were examined by a flow cytometer (FACSCantoII, BD Biosciences). (3) Measurement of NKT Cell Activating Action (IFN-γ Production) Any of human-derived CD14 positive cell lines (THP-1, U937, and human-derived iPS-CD14 positive cell were used here) and NKT cell ligand RK-163 were cultured for 8-48 hr in the presence or absence of GM-CSF to prepare “RK-163-pulsed human CD14 positive cell line”. ..

    Staining:

    Article Title: BK Polyomavirus-Specific CD8 T-Cell Expansion In Vitro Using 27mer Peptide Antigens for Developing Adoptive T-Cell Transfer and Vaccination.
    Article Snippet: Cells incubated with RPMI medium alone served as negative control and cells stimulated with staphylococcus enterotoxin B (3 μg/mL, Sigma-Aldrich, S4881) served as positive control. .. D ow nloaded from https://academ ic.oup.com /jid/article/223/8/1410/5898575 by guest on 08 M ay 2024 1412 • jid 2021:223 (15 April) • Wilhelm et al For extracellular staining, the following antibodies were used: CD14 (BD, 555399), CD209 (Miltenyi Biotec, 130-099-707), HLA-DR (BioLegend, San Diego, California, 307617), CD86 (BioLegend, 305426), CD83 (BioLegend, 305326), CD4 (BD, 562424), CD8 (BioLegend, 301012), CCR7 (BD, 561144), CD45RA (BD, 550855), PD1 (BD, 557946), CD69 (BD, 555530), CD107a (BD, 561348), and phycoerythrin-labeled streptamer (IBA, Hamburg, Germany). .. For intracellular staining, the following antibodies were used: CD207 (BD, 564727), CD1a (BD, 563939), IFN-γ (BD, 341117), and TNF-α (BD, 554512).

    Article Title: The composition of the gut microbiota following early-life antibiotic exposure affects host health and longevity in later life.
    Article Snippet: Anti-mouse antibodies used for cell surface staining were CD19, NK1.1, B220, TCR-B, CD11b, CD11c, MHC2, CD8, CD44, CD73 all from BD; CD4 from Biolegend; and CD62L, Ly6G, CD209, CD279, TNFa, all from Miltenyi. .. Anti-mouse antibodies used for cell surface staining were CD19, NK1.1, B220, TCR-B, CD11b, CD11c, MHC2, CD8, CD44, CD73 all from BD; CD4 from Biolegend; and CD62L, Ly6G, CD209, CD279, TNFa, all from Miltenyi. .. Red blood cells were lysedwith BD Pharm Lyse (BD Biosciences, San Jose, USA).

    Streptamer:

    Article Title: BK Polyomavirus-Specific CD8 T-Cell Expansion In Vitro Using 27mer Peptide Antigens for Developing Adoptive T-Cell Transfer and Vaccination.
    Article Snippet: Cells incubated with RPMI medium alone served as negative control and cells stimulated with staphylococcus enterotoxin B (3 μg/mL, Sigma-Aldrich, S4881) served as positive control. .. D ow nloaded from https://academ ic.oup.com /jid/article/223/8/1410/5898575 by guest on 08 M ay 2024 1412 • jid 2021:223 (15 April) • Wilhelm et al For extracellular staining, the following antibodies were used: CD14 (BD, 555399), CD209 (Miltenyi Biotec, 130-099-707), HLA-DR (BioLegend, San Diego, California, 307617), CD86 (BioLegend, 305426), CD83 (BioLegend, 305326), CD4 (BD, 562424), CD8 (BioLegend, 301012), CCR7 (BD, 561144), CD45RA (BD, 550855), PD1 (BD, 557946), CD69 (BD, 555530), CD107a (BD, 561348), and phycoerythrin-labeled streptamer (IBA, Hamburg, Germany). .. For intracellular staining, the following antibodies were used: CD207 (BD, 564727), CD1a (BD, 563939), IFN-γ (BD, 341117), and TNF-α (BD, 554512).

    Magnetic Cell Separation:

    Article Title: Macrophage Differentiation and Polarization Regulate the Release of the Immune Checkpoint Protein V-Domain Ig Suppressor of T Cell Activation
    Article Snippet: .. For flow cytometric analysis, human monocyte-derived macrophages (M1 and M2) were blocked for 30 min at 4°C with 50% FBS in autoMACS Rinsing Solution supplemented with MACS BSA Stock Solution (Miltenyi, Germany) and incubated with fluorescently labeled antibodies directed against, CD68 (Clone: REA886, Miltenyi Germany), CD80 (Clone: REA661, Miltenyi Germany), CD209 (Clone: REA617, Miltenyi Germany), and monoclonal mouse IgG2B anti-human VISTA (Clone: # 730804, R&D Systems, USA) as well as corresponding isotype control antibodies for 30 min at 4°C. .. Dead cells were stained with Propidium Iodide Solution (Miltenyi Biotec, Germany).

    Incubation:

    Article Title: Macrophage Differentiation and Polarization Regulate the Release of the Immune Checkpoint Protein V-Domain Ig Suppressor of T Cell Activation
    Article Snippet: .. For flow cytometric analysis, human monocyte-derived macrophages (M1 and M2) were blocked for 30 min at 4°C with 50% FBS in autoMACS Rinsing Solution supplemented with MACS BSA Stock Solution (Miltenyi, Germany) and incubated with fluorescently labeled antibodies directed against, CD68 (Clone: REA886, Miltenyi Germany), CD80 (Clone: REA661, Miltenyi Germany), CD209 (Clone: REA617, Miltenyi Germany), and monoclonal mouse IgG2B anti-human VISTA (Clone: # 730804, R&D Systems, USA) as well as corresponding isotype control antibodies for 30 min at 4°C. .. Dead cells were stained with Propidium Iodide Solution (Miltenyi Biotec, Germany).

    Article Title: The characterization, cytotoxicity, macrophage response and tissue regeneration of decellularized cartilage in costal cartilage defects.
    Article Snippet: After harvesting multiple costal cartilages, the local defect disrupts the integrity of the chest wall and may lead to obvious thoracic complications, such as local depression and asymmetry of the bilateral thoracic height.. Decellularized materials have been used for tissue reconstruction in clinical surgeries.. To apply xenogenic decellularized cartilage in costal cartilage defects, porcine-derived auricular and costal cartilage was tested for characterization, cytotoxicity, macrophage response, and tissue regeneration.

    Labeling:

    Article Title: Macrophage Differentiation and Polarization Regulate the Release of the Immune Checkpoint Protein V-Domain Ig Suppressor of T Cell Activation
    Article Snippet: .. For flow cytometric analysis, human monocyte-derived macrophages (M1 and M2) were blocked for 30 min at 4°C with 50% FBS in autoMACS Rinsing Solution supplemented with MACS BSA Stock Solution (Miltenyi, Germany) and incubated with fluorescently labeled antibodies directed against, CD68 (Clone: REA886, Miltenyi Germany), CD80 (Clone: REA661, Miltenyi Germany), CD209 (Clone: REA617, Miltenyi Germany), and monoclonal mouse IgG2B anti-human VISTA (Clone: # 730804, R&D Systems, USA) as well as corresponding isotype control antibodies for 30 min at 4°C. .. Dead cells were stained with Propidium Iodide Solution (Miltenyi Biotec, Germany).

    Control:

    Article Title: Macrophage Differentiation and Polarization Regulate the Release of the Immune Checkpoint Protein V-Domain Ig Suppressor of T Cell Activation
    Article Snippet: .. For flow cytometric analysis, human monocyte-derived macrophages (M1 and M2) were blocked for 30 min at 4°C with 50% FBS in autoMACS Rinsing Solution supplemented with MACS BSA Stock Solution (Miltenyi, Germany) and incubated with fluorescently labeled antibodies directed against, CD68 (Clone: REA886, Miltenyi Germany), CD80 (Clone: REA661, Miltenyi Germany), CD209 (Clone: REA617, Miltenyi Germany), and monoclonal mouse IgG2B anti-human VISTA (Clone: # 730804, R&D Systems, USA) as well as corresponding isotype control antibodies for 30 min at 4°C. .. Dead cells were stained with Propidium Iodide Solution (Miltenyi Biotec, Germany).

    other:




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