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cd14 microbeads  (Miltenyi Biotec)


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    Miltenyi Biotec cd14 microbeads
    Rosasina drives the pro‐inflammatory polarization of primary human macrophages. Primary human monocyte‐derived macrophages were generated by differentiation in M‐CSF and subsequently treated with increasing concentrations of the ROS‐inducing agent Rosasina (1, 10, and 100 nM). Following treatment, macrophage polarization was assessed by multiparameter flow cytometry using established M1‐and M2‐associated surface markers. Cells were gated on live, single CD45 + <t>/CD14</t> + macrophages prior to analysis. (A–F) Representative overlaid histograms and contour plots showing expression of M1‐associated markers. IFN‐γ (A, B) and MHC‐II (D, E) across treatment conditions. Quantification of M1 marker expression, presented as percentage of positive cells (C, F). (G–L) Representative overlaid histograms and contour plots showing the expression of M2‐associated markers. CD206 (G, H) and CD163) (J, K). Quantification of M2 marker expression (I, L) across treatment groups. (M) Cell viability following Rosasina exposure assessed by MTT assay to evaluate potential cytotoxic effects of treatment. Data represent independent experiments performed with primary human donors ( n = 4/group) in biological replicates. Quantified data are presented as mean ± SD. Statistical analysis was performed using ANOVA, with significance indicated as p < 0.05 compared with mock‐treated controls.
    Cd14 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 98/100, based on 4691 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd14+microbeads+human/CD14+MicroBeads%2C+human/pmc13650181-239-26-28
    Average 98 stars, based on 4691 article reviews
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    Images

    1) Product Images from "A Selective Pro‐Oxidant Drives Redox Reprogramming to Induce Ferroptosis and Immune Remodeling in Glioblastoma"

    Article Title: A Selective Pro‐Oxidant Drives Redox Reprogramming to Induce Ferroptosis and Immune Remodeling in Glioblastoma

    Journal: Smart Medicine

    doi: 10.1002/smmd.70058

    Rosasina drives the pro‐inflammatory polarization of primary human macrophages. Primary human monocyte‐derived macrophages were generated by differentiation in M‐CSF and subsequently treated with increasing concentrations of the ROS‐inducing agent Rosasina (1, 10, and 100 nM). Following treatment, macrophage polarization was assessed by multiparameter flow cytometry using established M1‐and M2‐associated surface markers. Cells were gated on live, single CD45 + /CD14 + macrophages prior to analysis. (A–F) Representative overlaid histograms and contour plots showing expression of M1‐associated markers. IFN‐γ (A, B) and MHC‐II (D, E) across treatment conditions. Quantification of M1 marker expression, presented as percentage of positive cells (C, F). (G–L) Representative overlaid histograms and contour plots showing the expression of M2‐associated markers. CD206 (G, H) and CD163) (J, K). Quantification of M2 marker expression (I, L) across treatment groups. (M) Cell viability following Rosasina exposure assessed by MTT assay to evaluate potential cytotoxic effects of treatment. Data represent independent experiments performed with primary human donors ( n = 4/group) in biological replicates. Quantified data are presented as mean ± SD. Statistical analysis was performed using ANOVA, with significance indicated as p < 0.05 compared with mock‐treated controls.
    Figure Legend Snippet: Rosasina drives the pro‐inflammatory polarization of primary human macrophages. Primary human monocyte‐derived macrophages were generated by differentiation in M‐CSF and subsequently treated with increasing concentrations of the ROS‐inducing agent Rosasina (1, 10, and 100 nM). Following treatment, macrophage polarization was assessed by multiparameter flow cytometry using established M1‐and M2‐associated surface markers. Cells were gated on live, single CD45 + /CD14 + macrophages prior to analysis. (A–F) Representative overlaid histograms and contour plots showing expression of M1‐associated markers. IFN‐γ (A, B) and MHC‐II (D, E) across treatment conditions. Quantification of M1 marker expression, presented as percentage of positive cells (C, F). (G–L) Representative overlaid histograms and contour plots showing the expression of M2‐associated markers. CD206 (G, H) and CD163) (J, K). Quantification of M2 marker expression (I, L) across treatment groups. (M) Cell viability following Rosasina exposure assessed by MTT assay to evaluate potential cytotoxic effects of treatment. Data represent independent experiments performed with primary human donors ( n = 4/group) in biological replicates. Quantified data are presented as mean ± SD. Statistical analysis was performed using ANOVA, with significance indicated as p < 0.05 compared with mock‐treated controls.

    Techniques Used: Derivative Assay, Generated, Flow Cytometry, Analysis, Expressing, Marker, MTT Assay

    Related Articles

    Isolation:

    Article Title: Establishment of a high-content compatible platform to assess effects of monocyte-derived factors on neural stem cell proliferation and differentiation.
    Article Snippet: .. Monocytes were then isolated from thawed PBMCs via positive magnetic activated cell sorting (MACS) using CD14 MicroBeads Human (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s instructions with the exception that half the bead concentration was used. ..

    Article Title: The gut microbiota protein BOC1 exhibits immune checkpoint inhibitor-like activity by inhibiting myeloid-derived suppressor cell differentiation
    Article Snippet: Finally, PBMCs were counted with a LUNA-FLTM fluorescence cell counter using Acridine Orange/Propidium Iodide Stain for viability assessment (Logos Biosystems by Aligned Genetics, Inc., Villeneuve d’Ascq, France). .. CD14 + and CD3 + cells were purified by positive selection using CD14 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) and CD3 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) magnetic isolation beads, following the manufacturer’s instructions. ..

    Article Title: Rejuvenation of peripheral immune cells attenuates Alzheimer's disease-like pathologies and behavioral deficits in a mouse model.
    Article Snippet: .. PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), mononuclear sections were collected, washed with phosphate-buffered saline (PBS) three times, and then used for monocytes isolation using CD14 MicroBeads Human (Miltenyi Biotec, Germany) and MS sorting columns (Miltenyi Biotec, Germany). ..

    Article Title: EB1010, a putative Christensenellaceae-derived microbial peptide, exhibits anti-inflammatory activity and therapeutic potential in ulcerative colitis
    Article Snippet: .. The CD14-negative fraction was obtained following CD14+ cells depletion using CD14 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) magnetic isolation beads, following the manufacturer's instructions. ..

    Article Title: Rejuvenation of peripheral immune cells attenuates Alzheimer’s disease-like pathologies and behavioral deficits in a mouse model
    Article Snippet: .. PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), mononuclear sections were collected, washed with phosphate-buffered saline (PBS) three times, and then used for monocytes isolation using CD14 MicroBeads Human (Miltenyi Biotec, Germany) and MS sorting columns (Miltenyi Biotec, Germany). ..

    Article Title: Reprogramming the melanoma and immunosuppressive myeloid cells with esomeprazole-loaded PLGA nanoparticles
    Article Snippet: The analysis of the cell cycle was performed by determining the DNA content distribution after propidium iodide staining, using a Cytoflex S and Kaluza software (both Beckman Coulter) according to. .. Monocytes were isolated from PBMC using the CD14 MicroBeads human (Miltenyi Biotec) and cultured at 37°C and 5% CO 2 atmosphere in RPMI 1640 medium with 10% FBS, L-Glutamine, penicillin and streptomycin in the presence of eNP or ESO-NP (same concentration as for melanoma studies) for 24 h prior to harvesting (patients’ MDSCs). ..

    FACS:

    Article Title: Establishment of a high-content compatible platform to assess effects of monocyte-derived factors on neural stem cell proliferation and differentiation.
    Article Snippet: .. Monocytes were then isolated from thawed PBMCs via positive magnetic activated cell sorting (MACS) using CD14 MicroBeads Human (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s instructions with the exception that half the bead concentration was used. ..

    Magnetic Cell Separation:

    Article Title: Establishment of a high-content compatible platform to assess effects of monocyte-derived factors on neural stem cell proliferation and differentiation.
    Article Snippet: .. Monocytes were then isolated from thawed PBMCs via positive magnetic activated cell sorting (MACS) using CD14 MicroBeads Human (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s instructions with the exception that half the bead concentration was used. ..

    Concentration Assay:

    Article Title: Establishment of a high-content compatible platform to assess effects of monocyte-derived factors on neural stem cell proliferation and differentiation.
    Article Snippet: .. Monocytes were then isolated from thawed PBMCs via positive magnetic activated cell sorting (MACS) using CD14 MicroBeads Human (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s instructions with the exception that half the bead concentration was used. ..

    Article Title: Reprogramming the melanoma and immunosuppressive myeloid cells with esomeprazole-loaded PLGA nanoparticles
    Article Snippet: The analysis of the cell cycle was performed by determining the DNA content distribution after propidium iodide staining, using a Cytoflex S and Kaluza software (both Beckman Coulter) according to. .. Monocytes were isolated from PBMC using the CD14 MicroBeads human (Miltenyi Biotec) and cultured at 37°C and 5% CO 2 atmosphere in RPMI 1640 medium with 10% FBS, L-Glutamine, penicillin and streptomycin in the presence of eNP or ESO-NP (same concentration as for melanoma studies) for 24 h prior to harvesting (patients’ MDSCs). ..

    Purification:

    Article Title: The gut microbiota protein BOC1 exhibits immune checkpoint inhibitor-like activity by inhibiting myeloid-derived suppressor cell differentiation
    Article Snippet: Finally, PBMCs were counted with a LUNA-FLTM fluorescence cell counter using Acridine Orange/Propidium Iodide Stain for viability assessment (Logos Biosystems by Aligned Genetics, Inc., Villeneuve d’Ascq, France). .. CD14 + and CD3 + cells were purified by positive selection using CD14 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) and CD3 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) magnetic isolation beads, following the manufacturer’s instructions. ..

    Selection:

    Article Title: The gut microbiota protein BOC1 exhibits immune checkpoint inhibitor-like activity by inhibiting myeloid-derived suppressor cell differentiation
    Article Snippet: Finally, PBMCs were counted with a LUNA-FLTM fluorescence cell counter using Acridine Orange/Propidium Iodide Stain for viability assessment (Logos Biosystems by Aligned Genetics, Inc., Villeneuve d’Ascq, France). .. CD14 + and CD3 + cells were purified by positive selection using CD14 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) and CD3 MicroBeads human (Miltenyi Biotec, Bergisch Gladbach, Germany) magnetic isolation beads, following the manufacturer’s instructions. ..

    Gradient Centrifugation:

    Article Title: Rejuvenation of peripheral immune cells attenuates Alzheimer's disease-like pathologies and behavioral deficits in a mouse model.
    Article Snippet: .. PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), mononuclear sections were collected, washed with phosphate-buffered saline (PBS) three times, and then used for monocytes isolation using CD14 MicroBeads Human (Miltenyi Biotec, Germany) and MS sorting columns (Miltenyi Biotec, Germany). ..

    Article Title: Rejuvenation of peripheral immune cells attenuates Alzheimer’s disease-like pathologies and behavioral deficits in a mouse model
    Article Snippet: .. PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), mononuclear sections were collected, washed with phosphate-buffered saline (PBS) three times, and then used for monocytes isolation using CD14 MicroBeads Human (Miltenyi Biotec, Germany) and MS sorting columns (Miltenyi Biotec, Germany). ..

    Saline:

    Article Title: Rejuvenation of peripheral immune cells attenuates Alzheimer's disease-like pathologies and behavioral deficits in a mouse model.
    Article Snippet: .. PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), mononuclear sections were collected, washed with phosphate-buffered saline (PBS) three times, and then used for monocytes isolation using CD14 MicroBeads Human (Miltenyi Biotec, Germany) and MS sorting columns (Miltenyi Biotec, Germany). ..

    Article Title: Rejuvenation of peripheral immune cells attenuates Alzheimer’s disease-like pathologies and behavioral deficits in a mouse model
    Article Snippet: .. PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque (TBD Science, Tianjin, China), mononuclear sections were collected, washed with phosphate-buffered saline (PBS) three times, and then used for monocytes isolation using CD14 MicroBeads Human (Miltenyi Biotec, Germany) and MS sorting columns (Miltenyi Biotec, Germany). ..

    Incubation:

    Article Title: Impact of cooperative or competitive dynamics between the yeast Saccharomyces cerevisiae and lactobacilli on the immune response of the host.
    Article Snippet: .. 100 ml of CD14 MicroBeads Human (Miltenyi Biotec) were added to the remaining pellet and incubated at 4°C for 15 minutes. .. Following the addition of 10 mL of MACS Buffer, CD14+ monocytes were isolated by positive magnetic separation using LS columns and the MidiMACSTM Separator immunomagnetic separator, according to the manufacturer’s protocol (Miltenyi Biotec).

    Article Title: Impact of cooperative or competitive dynamics between the yeast Saccharomyces cerevisiae and lactobacilli on the immune response of the host
    Article Snippet: .. 100 μl of CD14 MicroBeads Human (Miltenyi Biotec) were added to the remaining pellet and incubated at 4°C for 15 minutes. .. Following the addition of 10 mL of MACS Buffer, CD14+ monocytes were isolated by positive magnetic separation using LS columns and the MidiMACSTM Separator immunomagnetic separator, according to the manufacturer’s protocol (Miltenyi Biotec).

    Cell Culture:

    Article Title: Reprogramming the melanoma and immunosuppressive myeloid cells with esomeprazole-loaded PLGA nanoparticles
    Article Snippet: The analysis of the cell cycle was performed by determining the DNA content distribution after propidium iodide staining, using a Cytoflex S and Kaluza software (both Beckman Coulter) according to. .. Monocytes were isolated from PBMC using the CD14 MicroBeads human (Miltenyi Biotec) and cultured at 37°C and 5% CO 2 atmosphere in RPMI 1640 medium with 10% FBS, L-Glutamine, penicillin and streptomycin in the presence of eNP or ESO-NP (same concentration as for melanoma studies) for 24 h prior to harvesting (patients’ MDSCs). ..



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    Rosasina drives the pro‐inflammatory polarization of primary human macrophages. Primary human monocyte‐derived macrophages were generated by differentiation in M‐CSF and subsequently treated with increasing concentrations of the ROS‐inducing agent Rosasina (1, 10, and 100 nM). Following treatment, macrophage polarization was assessed by multiparameter flow cytometry using established M1‐and M2‐associated surface markers. Cells were gated on live, single CD45 + /CD14 + macrophages prior to analysis. (A–F) Representative overlaid histograms and contour plots showing expression of M1‐associated markers. IFN‐γ (A, B) and MHC‐II (D, E) across treatment conditions. Quantification of M1 marker expression, presented as percentage of positive cells (C, F). (G–L) Representative overlaid histograms and contour plots showing the expression of M2‐associated markers. CD206 (G, H) and CD163) (J, K). Quantification of M2 marker expression (I, L) across treatment groups. (M) Cell viability following Rosasina exposure assessed by MTT assay to evaluate potential cytotoxic effects of treatment. Data represent independent experiments performed with primary human donors ( n = 4/group) in biological replicates. Quantified data are presented as mean ± SD. Statistical analysis was performed using ANOVA, with significance indicated as p < 0.05 compared with mock‐treated controls.

    Journal: Smart Medicine

    Article Title: A Selective Pro‐Oxidant Drives Redox Reprogramming to Induce Ferroptosis and Immune Remodeling in Glioblastoma

    doi: 10.1002/smmd.70058

    Figure Lengend Snippet: Rosasina drives the pro‐inflammatory polarization of primary human macrophages. Primary human monocyte‐derived macrophages were generated by differentiation in M‐CSF and subsequently treated with increasing concentrations of the ROS‐inducing agent Rosasina (1, 10, and 100 nM). Following treatment, macrophage polarization was assessed by multiparameter flow cytometry using established M1‐and M2‐associated surface markers. Cells were gated on live, single CD45 + /CD14 + macrophages prior to analysis. (A–F) Representative overlaid histograms and contour plots showing expression of M1‐associated markers. IFN‐γ (A, B) and MHC‐II (D, E) across treatment conditions. Quantification of M1 marker expression, presented as percentage of positive cells (C, F). (G–L) Representative overlaid histograms and contour plots showing the expression of M2‐associated markers. CD206 (G, H) and CD163) (J, K). Quantification of M2 marker expression (I, L) across treatment groups. (M) Cell viability following Rosasina exposure assessed by MTT assay to evaluate potential cytotoxic effects of treatment. Data represent independent experiments performed with primary human donors ( n = 4/group) in biological replicates. Quantified data are presented as mean ± SD. Statistical analysis was performed using ANOVA, with significance indicated as p < 0.05 compared with mock‐treated controls.

    Article Snippet: CD14 + monocytes were isolated from peripheral blood mononuclear cells (PBMCs) using Ficoll‐Paque (GE Healthcare, Piscataway, NJ, USA) density‐gradient centrifugation followed by magnetic bead selection with CD14 MicroBeads (Miltenyi Biotec, Cologne, Germany) according to the manufacturer's protocol.

    Techniques: Derivative Assay, Generated, Flow Cytometry, Analysis, Expressing, Marker, MTT Assay