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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
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1r 8 s 9 s bicyclo 6 1 0 non 4 yn 9 ylmethyloxycarbonyl 1 8 diamino 3 6 dioxaoctane bcn  (Lifecore Biomedical)
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
1r 8 S 9 S Bicyclo 6 1 0 Non 4 Yn 9 Ylmethyloxycarbonyl 1 8 Diamino 3 6 Dioxaoctane Bcn, supplied by Lifecore Biomedical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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blz945 4 2 1r 2 r 2 hydroxycyclohexylamino benzothiazol 6 yloxyl pyridine 2 carboxylic acid methylamide  (Novartis)
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Evaluation of the <t>HEK-Blue</t> <t>IL-1R</t> reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.
Blz945 4 2 1r 2 R 2 Hydroxycyclohexylamino Benzothiazol 6 Yloxyl Pyridine 2 Carboxylic Acid Methylamide, supplied by Novartis, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Evaluation of the HEK-Blue IL-1R reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.

Journal: Molecules

Article Title: Cell Reporter-Based Analysis of Anti-Inflammasome Activities of Flavonoids with Distinct Structures

doi: 10.3390/molecules31183176

Figure Lengend Snippet: Evaluation of the HEK-Blue IL-1R reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.

Article Snippet: HEK-Blue IL-1R reporter cells were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Comparison, Generated, Activity Assay, Incubation

Application of the IL-1R HEK-Blue platform to compare the effects of flavonoids. ( A ) Experimental workflow for the detection of IL-1β in supernatants from BMMs treated with flavonoids and/or LPS using IL-1R HEK-Blue cells. ( B ) Comparison of SEAP activity, measured as OD at 640 nm, in Hek-Blue supernatants following exposure to media from BMMs cultured with or without LPS (10 ng/mL, 24 h). ( C ) Effects of flavonoid pretreatment (5–50 µM; NOB, QUER, or RUT) on SEAP induction by supernatants from LPS-treated BMMs. The −LPS and +LPS dashed lines indicate the mean OD values at 10 min from panel ( B ). ( D ) Comparison of OD values in Hek-Blue cell supernatants following exposure to supernatants from BMMs pretreated with flavonoids (10 µM). N = 12 for the −LPS and + LPS treatment groups and n = 4 for the flavonoid pretreatment groups. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Journal: Molecules

Article Title: Cell Reporter-Based Analysis of Anti-Inflammasome Activities of Flavonoids with Distinct Structures

doi: 10.3390/molecules31183176

Figure Lengend Snippet: Application of the IL-1R HEK-Blue platform to compare the effects of flavonoids. ( A ) Experimental workflow for the detection of IL-1β in supernatants from BMMs treated with flavonoids and/or LPS using IL-1R HEK-Blue cells. ( B ) Comparison of SEAP activity, measured as OD at 640 nm, in Hek-Blue supernatants following exposure to media from BMMs cultured with or without LPS (10 ng/mL, 24 h). ( C ) Effects of flavonoid pretreatment (5–50 µM; NOB, QUER, or RUT) on SEAP induction by supernatants from LPS-treated BMMs. The −LPS and +LPS dashed lines indicate the mean OD values at 10 min from panel ( B ). ( D ) Comparison of OD values in Hek-Blue cell supernatants following exposure to supernatants from BMMs pretreated with flavonoids (10 µM). N = 12 for the −LPS and + LPS treatment groups and n = 4 for the flavonoid pretreatment groups. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: HEK-Blue IL-1R reporter cells were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Comparison, Activity Assay, Cell Culture