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active cd73 enzyme (R&D Systems)

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R&D Systems active cd73 enzyme

A) Enzyme reaction for cN-II (Human cytosolic 5'-Nucleotidase) was carried out using increasing concentrations of enzyme and 10μM AMP for 60min at 37°C and activity was determined following AMP-Glo assay as described in Materials and Methods section. The results are shown as net RLU after subtraction of no enzyme control. The experiment was done in triplicates; results shown are mean ± SD. B) Selectivity of the inhibitor AMP-CP against <t>CD73</t> and cytosolic cN-II was determined using purified CD73 (0.1 ng/Rx) and cN-II (0.6 mU/Rx) by AMP-PC using 10μM AMP substrate at 23°C for 5 min (CD73) and at 37°C for 30 min (cN-II). Activities were determined following AMP-Glo protocol. The inhibitor inhibited CD73 activity with an IC 50 value of 3x10 -7 M with minimal or no inhibition of cN-II. Experiments were done in triplicates; results shown are mean ± SD. C) Monitoring enzymatic activity of membrane-associated CD73 in cell-based assay.

Active Cd73 Enzyme, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/active cd73 enzyme/product/R&D Systems
Average 94 stars, based on 14 article reviews

active cd73 enzyme - by Bioz Stars, 2026-03

94/100 stars

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1) Product Images from "Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39"

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

Journal: PLoS ONE

doi: 10.1371/journal.pone.0220094

Figure Legend Snippet: A) Enzyme reaction for cN-II (Human cytosolic 5'-Nucleotidase) was carried out using increasing concentrations of enzyme and 10μM AMP for 60min at 37°C and activity was determined following AMP-Glo assay as described in Materials and Methods section. The results are shown as net RLU after subtraction of no enzyme control. The experiment was done in triplicates; results shown are mean ± SD. B) Selectivity of the inhibitor AMP-CP against CD73 and cytosolic cN-II was determined using purified CD73 (0.1 ng/Rx) and cN-II (0.6 mU/Rx) by AMP-PC using 10μM AMP substrate at 23°C for 5 min (CD73) and at 37°C for 30 min (cN-II). Activities were determined following AMP-Glo protocol. The inhibitor inhibited CD73 activity with an IC 50 value of 3x10 -7 M with minimal or no inhibition of cN-II. Experiments were done in triplicates; results shown are mean ± SD. C) Monitoring enzymatic activity of membrane-associated CD73 in cell-based assay.

Techniques Used: Activity Assay, Glo Assay, Control, Purification, Inhibition, Membrane, Cell Based Assay

A) Five cell lines were evaluated for their CD73 enzyme activity in intact cells using AMP Glo protocol for cells as described in the method section. Time course of enzyme activity with 10 μM AMP following AMP-Glo protocol. Cells: T-47D (circle), MDA-MB-231(square), SK-MEL2 (triangle), A375 (reverse triangle), and SK-OV3 (diamond). B) Determination of abundance of CD73 in membranes of five cell lines using western blotting. Cell lysates (10μg) from each cell line and pure CD73 (0.2μg) as positive control (lane 7) were run on gels and immunoblotted using primary antibodies (anti- NT5E/CD73, Cell Signaling), incubated overnight at 4°C followed by HRP-ECL as described in the method section. C) Membrane associated CD73 bound MDA-MB-231 activity was determined in presence (open circle) and absence (closed circle) of 50μM AMP-PC. Reactions contained 25k MDA-MB-231 cell per well and incubated at 37°C with 10μM AMP in final 250μl per well. Aliquots of 25μl per sample were withdrawn at each time point and activity was determined following AMP-Glo assay. Each point represents the average of triplicates; the error bars represent the SD.

Figure Legend Snippet: A) Five cell lines were evaluated for their CD73 enzyme activity in intact cells using AMP Glo protocol for cells as described in the method section. Time course of enzyme activity with 10 μM AMP following AMP-Glo protocol. Cells: T-47D (circle), MDA-MB-231(square), SK-MEL2 (triangle), A375 (reverse triangle), and SK-OV3 (diamond). B) Determination of abundance of CD73 in membranes of five cell lines using western blotting. Cell lysates (10μg) from each cell line and pure CD73 (0.2μg) as positive control (lane 7) were run on gels and immunoblotted using primary antibodies (anti- NT5E/CD73, Cell Signaling), incubated overnight at 4°C followed by HRP-ECL as described in the method section. C) Membrane associated CD73 bound MDA-MB-231 activity was determined in presence (open circle) and absence (closed circle) of 50μM AMP-PC. Reactions contained 25k MDA-MB-231 cell per well and incubated at 37°C with 10μM AMP in final 250μl per well. Aliquots of 25μl per sample were withdrawn at each time point and activity was determined following AMP-Glo assay. Each point represents the average of triplicates; the error bars represent the SD.

Techniques Used: Activity Assay, Western Blot, Positive Control, Incubation, Membrane, Glo Assay

Selectivity of various inhibitors against soluble CD73 and membrane bound CD73 enzyme using assay protocol for purified and cell-based assays.

Figure Legend Snippet: Selectivity of various inhibitors against soluble CD73 and membrane bound CD73 enzyme using assay protocol for purified and cell-based assays.

Techniques Used: Membrane, Purification

Figure Legend Snippet: Effect of various anti-CD73 antibodies on blocking the activity of CD73 bound MDA-MB-231 cells.

Techniques Used: Blocking Assay, Activity Assay

Inhibitors of CD39 and CD73 were incubated with purified CD39 (A, B) and Farage associated enzyme (C, D) and assayed for CD39 activity using ATP (A, C) or ADP (B, D) as substrate. Results show percentage inhibition of CD39 activity using ARL 67156 (circle), POM1 (solid square), and AMP-CP (triangle). Purified CD39 (0.1ng) and 10μM ATP or ADP in each reaction for 30min at 37°C; for cell-based Farage cells (200K cells) were incubated with 10μM ATP or ADP in each reaction for 20min at 37°C. Experiments were carried out in triplicates; results shown are mean ± SD.

Figure Legend Snippet: Inhibitors of CD39 and CD73 were incubated with purified CD39 (A, B) and Farage associated enzyme (C, D) and assayed for CD39 activity using ATP (A, C) or ADP (B, D) as substrate. Results show percentage inhibition of CD39 activity using ARL 67156 (circle), POM1 (solid square), and AMP-CP (triangle). Purified CD39 (0.1ng) and 10μM ATP or ADP in each reaction for 30min at 37°C; for cell-based Farage cells (200K cells) were incubated with 10μM ATP or ADP in each reaction for 20min at 37°C. Experiments were carried out in triplicates; results shown are mean ± SD.

Techniques Used: Incubation, Purification, Activity Assay, Inhibition

Testing the specificity of CD39 inhibitor POM1 (A) and CD73 inhibitor AMP-CP (B) on the activity of different kinases, ATPase, as well as CD39 and CD73. Enzyme tested are protein serine/threonine/tyrosine, lipid, sugar, and inorganic kinases, and K/Na ATPase. Results show that the majority of enzymes are inhibited by POM1 (A), but the more selective CD73 inhibitor AMP-PC) was highly selective for CD73 with minimal inhibition for PKA and no inhibition of the other enzymes. PKA, protein kinase A (solid circle), PKCα (solid square), Src, protein tyrosine Src kinase (solid triangle), Acka, acetate kinase from Escherichia coli (solid reverse triangle), HK, hexokinase from Saccharomyces cerevisiae (solid circle), PI3αKinase, p110α/p85α (square), K/K ATPase, Adenosine 5´-Triphosphatase from porcine cerebral cortex (circle), CD39 (ATP substrate, triangle), and CD39 (ADP substrate, reverse triangle); for CD73 (triangle) in panel B. Each point represents average of a typical experiment done in triplicates; results shown are mean ± SD.

Figure Legend Snippet: Testing the specificity of CD39 inhibitor POM1 (A) and CD73 inhibitor AMP-CP (B) on the activity of different kinases, ATPase, as well as CD39 and CD73. Enzyme tested are protein serine/threonine/tyrosine, lipid, sugar, and inorganic kinases, and K/Na ATPase. Results show that the majority of enzymes are inhibited by POM1 (A), but the more selective CD73 inhibitor AMP-PC) was highly selective for CD73 with minimal inhibition for PKA and no inhibition of the other enzymes. PKA, protein kinase A (solid circle), PKCα (solid square), Src, protein tyrosine Src kinase (solid triangle), Acka, acetate kinase from Escherichia coli (solid reverse triangle), HK, hexokinase from Saccharomyces cerevisiae (solid circle), PI3αKinase, p110α/p85α (square), K/K ATPase, Adenosine 5´-Triphosphatase from porcine cerebral cortex (circle), CD39 (ATP substrate, triangle), and CD39 (ADP substrate, reverse triangle); for CD73 (triangle) in panel B. Each point represents average of a typical experiment done in triplicates; results shown are mean ± SD.

Techniques Used: Activity Assay, Inhibition

Image Search Results

Journal: PLoS ONE

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

doi: 10.1371/journal.pone.0220094

Figure Lengend Snippet: A) Enzyme reaction for cN-II (Human cytosolic 5'-Nucleotidase) was carried out using increasing concentrations of enzyme and 10μM AMP for 60min at 37°C and activity was determined following AMP-Glo assay as described in Materials and Methods section. The results are shown as net RLU after subtraction of no enzyme control. The experiment was done in triplicates; results shown are mean ± SD. B) Selectivity of the inhibitor AMP-CP against CD73 and cytosolic cN-II was determined using purified CD73 (0.1 ng/Rx) and cN-II (0.6 mU/Rx) by AMP-PC using 10μM AMP substrate at 23°C for 5 min (CD73) and at 37°C for 30 min (cN-II). Activities were determined following AMP-Glo protocol. The inhibitor inhibited CD73 activity with an IC 50 value of 3x10 -7 M with minimal or no inhibition of cN-II. Experiments were done in triplicates; results shown are mean ± SD. C) Monitoring enzymatic activity of membrane-associated CD73 in cell-based assay.

Article Snippet: Active CD73 enzyme (Recombinant Human 5'-Nucleotidase/CD73 Protein, CF) and active CD39 (His-Tag Human) were from R&D Systems, Minneapolis, MN).

Techniques: Activity Assay, Glo Assay, Control, Purification, Inhibition, Membrane, Cell Based Assay

Journal: PLoS ONE

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

doi: 10.1371/journal.pone.0220094

Figure Lengend Snippet: A) Five cell lines were evaluated for their CD73 enzyme activity in intact cells using AMP Glo protocol for cells as described in the method section. Time course of enzyme activity with 10 μM AMP following AMP-Glo protocol. Cells: T-47D (circle), MDA-MB-231(square), SK-MEL2 (triangle), A375 (reverse triangle), and SK-OV3 (diamond). B) Determination of abundance of CD73 in membranes of five cell lines using western blotting. Cell lysates (10μg) from each cell line and pure CD73 (0.2μg) as positive control (lane 7) were run on gels and immunoblotted using primary antibodies (anti- NT5E/CD73, Cell Signaling), incubated overnight at 4°C followed by HRP-ECL as described in the method section. C) Membrane associated CD73 bound MDA-MB-231 activity was determined in presence (open circle) and absence (closed circle) of 50μM AMP-PC. Reactions contained 25k MDA-MB-231 cell per well and incubated at 37°C with 10μM AMP in final 250μl per well. Aliquots of 25μl per sample were withdrawn at each time point and activity was determined following AMP-Glo assay. Each point represents the average of triplicates; the error bars represent the SD.

Article Snippet: Active CD73 enzyme (Recombinant Human 5'-Nucleotidase/CD73 Protein, CF) and active CD39 (His-Tag Human) were from R&D Systems, Minneapolis, MN).

Techniques: Activity Assay, Western Blot, Positive Control, Incubation, Membrane, Glo Assay

Journal: PLoS ONE

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

doi: 10.1371/journal.pone.0220094

Figure Lengend Snippet: Selectivity of various inhibitors against soluble CD73 and membrane bound CD73 enzyme using assay protocol for purified and cell-based assays.

Article Snippet: Active CD73 enzyme (Recombinant Human 5'-Nucleotidase/CD73 Protein, CF) and active CD39 (His-Tag Human) were from R&D Systems, Minneapolis, MN).

Techniques: Membrane, Purification

Journal: PLoS ONE

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

doi: 10.1371/journal.pone.0220094

Figure Lengend Snippet: Effect of various anti-CD73 antibodies on blocking the activity of CD73 bound MDA-MB-231 cells.

Article Snippet: Active CD73 enzyme (Recombinant Human 5'-Nucleotidase/CD73 Protein, CF) and active CD39 (His-Tag Human) were from R&D Systems, Minneapolis, MN).

Techniques: Blocking Assay, Activity Assay

Journal: PLoS ONE

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

doi: 10.1371/journal.pone.0220094

Figure Lengend Snippet: Inhibitors of CD39 and CD73 were incubated with purified CD39 (A, B) and Farage associated enzyme (C, D) and assayed for CD39 activity using ATP (A, C) or ADP (B, D) as substrate. Results show percentage inhibition of CD39 activity using ARL 67156 (circle), POM1 (solid square), and AMP-CP (triangle). Purified CD39 (0.1ng) and 10μM ATP or ADP in each reaction for 30min at 37°C; for cell-based Farage cells (200K cells) were incubated with 10μM ATP or ADP in each reaction for 20min at 37°C. Experiments were carried out in triplicates; results shown are mean ± SD.

Article Snippet: Active CD73 enzyme (Recombinant Human 5'-Nucleotidase/CD73 Protein, CF) and active CD39 (His-Tag Human) were from R&D Systems, Minneapolis, MN).

Techniques: Incubation, Purification, Activity Assay, Inhibition

Journal: PLoS ONE

Article Title: Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39

doi: 10.1371/journal.pone.0220094

Figure Lengend Snippet: Testing the specificity of CD39 inhibitor POM1 (A) and CD73 inhibitor AMP-CP (B) on the activity of different kinases, ATPase, as well as CD39 and CD73. Enzyme tested are protein serine/threonine/tyrosine, lipid, sugar, and inorganic kinases, and K/Na ATPase. Results show that the majority of enzymes are inhibited by POM1 (A), but the more selective CD73 inhibitor AMP-PC) was highly selective for CD73 with minimal inhibition for PKA and no inhibition of the other enzymes. PKA, protein kinase A (solid circle), PKCα (solid square), Src, protein tyrosine Src kinase (solid triangle), Acka, acetate kinase from Escherichia coli (solid reverse triangle), HK, hexokinase from Saccharomyces cerevisiae (solid circle), PI3αKinase, p110α/p85α (square), K/K ATPase, Adenosine 5´-Triphosphatase from porcine cerebral cortex (circle), CD39 (ATP substrate, triangle), and CD39 (ADP substrate, reverse triangle); for CD73 (triangle) in panel B. Each point represents average of a typical experiment done in triplicates; results shown are mean ± SD.

Article Snippet: Active CD73 enzyme (Recombinant Human 5'-Nucleotidase/CD73 Protein, CF) and active CD39 (His-Tag Human) were from R&D Systems, Minneapolis, MN).

Techniques: Activity Assay, Inhibition

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1) Product Images from "Monitoring and characterizing soluble and membrane-bound ectonucleotidases CD73 and CD39"

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