Structured Review

Millipore ampcp
<t>CD73</t> on macrophages or apoptotic cells can mediate TNF suppression during efferocytosis. ( a ) Above, flow cytometry analysis of CD39 and CD73 expression on surface of parental Jurkat cells or stable Jurkat cell lines expressing CD39 or CD73. Below, CD73 −/− LEC-MФ were stimulated with 100 ng/ml LPS±live or apoptotic Jurkat cells for 4 h in the absence or presence of 20 μ M <t>AMPCP</t> and TNF levels determined by ELISA. Mean±S.E.M. of three independent experiments shown. ( b ) RAW264.7 cells were transfected with plasmids encoding YFP or A2a-YFP and subsequently FACS-sorted by YFP expression. YFP + cells were plated and stimulated with LPS±apoptotic parental Jurkat cells or CD73-expressing Jurkat cells and TNF levels measured by ELISA. Mean±S.E.M. of three independent experiments shown. (* P
Ampcp, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ampcp - by Bioz Stars, 2020-09
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1) Product Images from "CD73 regulates anti-inflammatory signaling between apoptotic cells and endotoxin-conditioned tissue macrophages"

Article Title: CD73 regulates anti-inflammatory signaling between apoptotic cells and endotoxin-conditioned tissue macrophages

Journal: Cell Death and Differentiation

doi: 10.1038/cdd.2016.159

CD73 on macrophages or apoptotic cells can mediate TNF suppression during efferocytosis. ( a ) Above, flow cytometry analysis of CD39 and CD73 expression on surface of parental Jurkat cells or stable Jurkat cell lines expressing CD39 or CD73. Below, CD73 −/− LEC-MФ were stimulated with 100 ng/ml LPS±live or apoptotic Jurkat cells for 4 h in the absence or presence of 20 μ M AMPCP and TNF levels determined by ELISA. Mean±S.E.M. of three independent experiments shown. ( b ) RAW264.7 cells were transfected with plasmids encoding YFP or A2a-YFP and subsequently FACS-sorted by YFP expression. YFP + cells were plated and stimulated with LPS±apoptotic parental Jurkat cells or CD73-expressing Jurkat cells and TNF levels measured by ELISA. Mean±S.E.M. of three independent experiments shown. (* P
Figure Legend Snippet: CD73 on macrophages or apoptotic cells can mediate TNF suppression during efferocytosis. ( a ) Above, flow cytometry analysis of CD39 and CD73 expression on surface of parental Jurkat cells or stable Jurkat cell lines expressing CD39 or CD73. Below, CD73 −/− LEC-MФ were stimulated with 100 ng/ml LPS±live or apoptotic Jurkat cells for 4 h in the absence or presence of 20 μ M AMPCP and TNF levels determined by ELISA. Mean±S.E.M. of three independent experiments shown. ( b ) RAW264.7 cells were transfected with plasmids encoding YFP or A2a-YFP and subsequently FACS-sorted by YFP expression. YFP + cells were plated and stimulated with LPS±apoptotic parental Jurkat cells or CD73-expressing Jurkat cells and TNF levels measured by ELISA. Mean±S.E.M. of three independent experiments shown. (* P

Techniques Used: Flow Cytometry, Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Transfection, FACS

2) Product Images from "CD73 Activity is Dispensable for the Polarization of M2 Macrophages"

Article Title: CD73 Activity is Dispensable for the Polarization of M2 Macrophages

Journal: PLoS ONE

doi: 10.1371/journal.pone.0134721

CD73 activity is dispensable for polarization of human monocytes/macrophages. Purified human monocytes were polarized with LPS+TNF or IL-4+M-CSF in the presence or absence of CD73 inhibitor AMPCP for 3 days. (A) Flow cytometric analyses of CD14 and CD206 surface expression. (B) qPCR analyses of CCL19 and MRC1 expression. Results are shown as boxplots with 5–95 percentiles (n = 8 different donors) from 4 different experiments. None of the differences between the AMPCP-treated and control cells were statistically significant under any condition.
Figure Legend Snippet: CD73 activity is dispensable for polarization of human monocytes/macrophages. Purified human monocytes were polarized with LPS+TNF or IL-4+M-CSF in the presence or absence of CD73 inhibitor AMPCP for 3 days. (A) Flow cytometric analyses of CD14 and CD206 surface expression. (B) qPCR analyses of CCL19 and MRC1 expression. Results are shown as boxplots with 5–95 percentiles (n = 8 different donors) from 4 different experiments. None of the differences between the AMPCP-treated and control cells were statistically significant under any condition.

Techniques Used: Activity Assay, Purification, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

Induction of CD73 on M(LPS+TNF)-polarized human macrophages. (A) The expression of cell-surface CD73 on peripheral blood mononuclear cells and macrophages and the purity of the macrophage population after MACS-selection is shown. Ficoll-isolated cells were analyzed by flow cytometry before and immediately after MACS selection for CD14 (a monocyte marker) and CD73 expression and for forward (FSC) and side (SSC) scatters. The percentages of cells falling into different quadrants are indicated. (B) The average CD73 expression on CD14 + monocytes polarized with and without 100 μM AMPCP to M(LPS+TNF) or M(IL-4+M-CSF) cells after 3 days. Box plots and representative dot plots for each polarization are shown. (C) Expression of NT5E (= CD73) on the mRNA level after the indicated polarizations is shown as deltaCT values. (D) Enzymatic activity of 5’-ectonucleotidase (= CD73) after the indicated polarization. In (B), (C) and (D) the vertical line represents the median and the whiskers the 5–95 percentiles. Pooled data (n = 7–8 different donors) from at least 2 different experiments are shown.
Figure Legend Snippet: Induction of CD73 on M(LPS+TNF)-polarized human macrophages. (A) The expression of cell-surface CD73 on peripheral blood mononuclear cells and macrophages and the purity of the macrophage population after MACS-selection is shown. Ficoll-isolated cells were analyzed by flow cytometry before and immediately after MACS selection for CD14 (a monocyte marker) and CD73 expression and for forward (FSC) and side (SSC) scatters. The percentages of cells falling into different quadrants are indicated. (B) The average CD73 expression on CD14 + monocytes polarized with and without 100 μM AMPCP to M(LPS+TNF) or M(IL-4+M-CSF) cells after 3 days. Box plots and representative dot plots for each polarization are shown. (C) Expression of NT5E (= CD73) on the mRNA level after the indicated polarizations is shown as deltaCT values. (D) Enzymatic activity of 5’-ectonucleotidase (= CD73) after the indicated polarization. In (B), (C) and (D) the vertical line represents the median and the whiskers the 5–95 percentiles. Pooled data (n = 7–8 different donors) from at least 2 different experiments are shown.

Techniques Used: Expressing, Magnetic Cell Separation, Selection, Isolation, Flow Cytometry, Cytometry, Marker, Activity Assay

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Article Snippet: Adenosine 5′-[γ-thio]triphosphate tetralithium salt (ATP-γ-S), α,β-methyleneadenosine 5′-triphosphate lithium salt (α,β-Me-ATP), BzATP, α,β-methyleneadenosine 5′-diphosphate sodium salt (α,β-Me-ADP), and 2-methylthioadenosine diphosphate trisodium salt (2-MeS-ADP) were purchased from Sigma–Aldrich.

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Article Title: Adenosine A3 receptor elicits chemoresistance mediated by multiple resistance-associated protein-1 in human glioblastoma stem-like cells
Article Snippet: .. Pharmacological agents For in vitro studies AOPCP (50 μM; α,β-Methyleneadenosine 5′-diphosphate) was used as a CD73 inhibitor (M8386; Sigma, Saint Louis, MO) and MRS1220 (#1217; 10 μM) as a selective antagonist of A3 AR [ ]. ..

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