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Bio-Rad pgas cis-reporter plasmids
$A, Measurements of FA value <t>of</t> <t>RAW</t> 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for varying time periods. B , RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for 18 hrs (time equivalent of total parasite-MØ interaction of 6 hrs of initial attachment and 12 hours of infection establishment) were subjected to membrane harvestation. Gradient fractions were purified as described in . Equal volume of fractions from each optiprep gradient were analyzed by dot blot for the presence of GM1, IFNγR1, LPG by using the appropriate antibodies and CTxB-HRP. C, IFNγ induced bioactivity in LPG treated MØs was determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven <t>pGAS</t> reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) and /or treated with 10 µg/ml of LPG for 18 hours. Treated cells were then replenished or not with CH-Lipo. Subsequently treated cells were stimulated with or without 100 U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. The data represents the relative luciferase activity normalized to per microgram of protein content. Data are representative of three independent experiments.$
Pgas Cis Reporter Plasmids, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pgas+cis-reporter+plasmids/pmc03169561-280-7-22?v=Bio-Rad
Average 90 stars, based on 1 article reviews

pgas cis-reporter plasmids - by Bioz Stars, 2026-07

90/100 stars

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1) Product Images from "Restoration of IFNγR Subunit Assembly, IFNγ Signaling and Parasite Clearance in Leishmania donovani Infected Macrophages: Role of Membrane Cholesterol"

Article Title: Restoration of IFNγR Subunit Assembly, IFNγ Signaling and Parasite Clearance in Leishmania donovani Infected Macrophages: Role of Membrane Cholesterol

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1002229

$A, Measurements of FA value of RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for varying time periods. B , RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for 18 hrs (time equivalent of total parasite-MØ interaction of 6 hrs of initial attachment and 12 hours of infection establishment) were subjected to membrane harvestation. Gradient fractions were purified as described in . Equal volume of fractions from each optiprep gradient were analyzed by dot blot for the presence of GM1, IFNγR1, LPG by using the appropriate antibodies and CTxB-HRP. C, IFNγ induced bioactivity in LPG treated MØs was determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven pGAS reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) and /or treated with 10 µg/ml of LPG for 18 hours. Treated cells were then replenished or not with CH-Lipo. Subsequently treated cells were stimulated with or without 100 U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. The data represents the relative luciferase activity normalized to per microgram of protein content. Data are representative of three independent experiments.$
Figure Legend Snippet: A, Measurements of FA value of RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for varying time periods. B , RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for 18 hrs (time equivalent of total parasite-MØ interaction of 6 hrs of initial attachment and 12 hours of infection establishment) were subjected to membrane harvestation. Gradient fractions were purified as described in . Equal volume of fractions from each optiprep gradient were analyzed by dot blot for the presence of GM1, IFNγR1, LPG by using the appropriate antibodies and CTxB-HRP. C, IFNγ induced bioactivity in LPG treated MØs was determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven pGAS reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) and /or treated with 10 µg/ml of LPG for 18 hours. Treated cells were then replenished or not with CH-Lipo. Subsequently treated cells were stimulated with or without 100 U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. The data represents the relative luciferase activity normalized to per microgram of protein content. Data are representative of three independent experiments.

Techniques Used: Purification, Infection, Dot Blot, Reporter Assay, Transfection, Negative Control, Plasmid Preparation, Luciferase, Activity Assay

Infected RAW 264.7 cells were left untreated or treated with CH-Lipo or AN-Lipo or DPPC-Lipo followed by stimulation with 100 U/ml of rIFNγ for 24 hours at 4 th hour post infection ( A, B, C ) or 12 th hour post infection ( A', B', C' ) and the number of intracellular amastigotes per MØ, % infected MØs and nitrite concentration (µM) were determined. Data are representative of three independent experiments. D , Pretreatment with inducible nitric oxide synthase inhibitor blocks IFNγ induced leishmanicidal effect in CH-Lipo treated infected MØs. RAW 264.7 cells were infected with LD at m.o.i of 10, for 6 hours. MØs were then washed and either treated with CH-Lipo or left untreated as infected control. Subsequently, at indicated timepoints postinfection, MØs were stimulated with or without 100 U/ml of rIFNγ for 24 hours in presence or absence of 5 µM L-NMMA. Cell free supernatant was harvested for NO measurement. Data of Nitrite concentration (µM) is representative of three independent experiments. The amastigote numbers of the parallel sets treated similarly was demonstrated in D' . E , Enhanced IFNγ-bioactivity in CH-Lipo replenished MØs as determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven pGAS reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) or infected with LD. Treated cells were then replenished or not with CH-Lipo. Subsequently, mBCD treated cells with or without CH-Lipo reloading were stimulated with or without 100U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. While the infected cells with or without CH-Lipo treatment were stimulated or not with 100 U/ml of rIFNγ at indicated timepoints post infection. Eight hours later, luciferase activities in cell lysates were measured as described in . The data represents the relative luciferase activity normalized to per microgram of protein content. Data is representative of three independent experiments.

Figure Legend Snippet: Infected RAW 264.7 cells were left untreated or treated with CH-Lipo or AN-Lipo or DPPC-Lipo followed by stimulation with 100 U/ml of rIFNγ for 24 hours at 4 th hour post infection ( A, B, C ) or 12 th hour post infection ( A', B', C' ) and the number of intracellular amastigotes per MØ, % infected MØs and nitrite concentration (µM) were determined. Data are representative of three independent experiments. D , Pretreatment with inducible nitric oxide synthase inhibitor blocks IFNγ induced leishmanicidal effect in CH-Lipo treated infected MØs. RAW 264.7 cells were infected with LD at m.o.i of 10, for 6 hours. MØs were then washed and either treated with CH-Lipo or left untreated as infected control. Subsequently, at indicated timepoints postinfection, MØs were stimulated with or without 100 U/ml of rIFNγ for 24 hours in presence or absence of 5 µM L-NMMA. Cell free supernatant was harvested for NO measurement. Data of Nitrite concentration (µM) is representative of three independent experiments. The amastigote numbers of the parallel sets treated similarly was demonstrated in D' . E , Enhanced IFNγ-bioactivity in CH-Lipo replenished MØs as determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven pGAS reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) or infected with LD. Treated cells were then replenished or not with CH-Lipo. Subsequently, mBCD treated cells with or without CH-Lipo reloading were stimulated with or without 100U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. While the infected cells with or without CH-Lipo treatment were stimulated or not with 100 U/ml of rIFNγ at indicated timepoints post infection. Eight hours later, luciferase activities in cell lysates were measured as described in . The data represents the relative luciferase activity normalized to per microgram of protein content. Data is representative of three independent experiments.

Techniques Used: Infection, Concentration Assay, Reporter Assay, Transfection, Negative Control, Plasmid Preparation, Luciferase, Activity Assay

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Journal: PLoS Pathogens

Article Title: Restoration of IFNγR Subunit Assembly, IFNγ Signaling and Parasite Clearance in Leishmania donovani Infected Macrophages: Role of Membrane Cholesterol

doi: 10.1371/journal.ppat.1002229

Figure Lengend Snippet: A, Measurements of FA value of RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for varying time periods. B , RAW 264.7 cells left untreated or treated with 10 µg/ml of purified LPG for 18 hrs (time equivalent of total parasite-MØ interaction of 6 hrs of initial attachment and 12 hours of infection establishment) were subjected to membrane harvestation. Gradient fractions were purified as described in . Equal volume of fractions from each optiprep gradient were analyzed by dot blot for the presence of GM1, IFNγR1, LPG by using the appropriate antibodies and CTxB-HRP. C, IFNγ induced bioactivity in LPG treated MØs was determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven pGAS reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) and /or treated with 10 µg/ml of LPG for 18 hours. Treated cells were then replenished or not with CH-Lipo. Subsequently treated cells were stimulated with or without 100 U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. The data represents the relative luciferase activity normalized to per microgram of protein content. Data are representative of three independent experiments.

Article Snippet: RAW 264.7 cells were transfected with the pGAS cis-reporter plasmids or the pCIS-CK negative control plasmid by electroporation by Genepulser X cell (Bio-Rad, Hercules, CA).

Techniques: Purification, Infection, Dot Blot, Reporter Assay, Transfection, Negative Control, Plasmid Preparation, Luciferase, Activity Assay

Journal: PLoS Pathogens

Article Title: Restoration of IFNγR Subunit Assembly, IFNγ Signaling and Parasite Clearance in Leishmania donovani Infected Macrophages: Role of Membrane Cholesterol

doi: 10.1371/journal.ppat.1002229

Figure Lengend Snippet: Infected RAW 264.7 cells were left untreated or treated with CH-Lipo or AN-Lipo or DPPC-Lipo followed by stimulation with 100 U/ml of rIFNγ for 24 hours at 4 th hour post infection ( A, B, C ) or 12 th hour post infection ( A', B', C' ) and the number of intracellular amastigotes per MØ, % infected MØs and nitrite concentration (µM) were determined. Data are representative of three independent experiments. D , Pretreatment with inducible nitric oxide synthase inhibitor blocks IFNγ induced leishmanicidal effect in CH-Lipo treated infected MØs. RAW 264.7 cells were infected with LD at m.o.i of 10, for 6 hours. MØs were then washed and either treated with CH-Lipo or left untreated as infected control. Subsequently, at indicated timepoints postinfection, MØs were stimulated with or without 100 U/ml of rIFNγ for 24 hours in presence or absence of 5 µM L-NMMA. Cell free supernatant was harvested for NO measurement. Data of Nitrite concentration (µM) is representative of three independent experiments. The amastigote numbers of the parallel sets treated similarly was demonstrated in D' . E , Enhanced IFNγ-bioactivity in CH-Lipo replenished MØs as determined by reporter assay. RAW 264.7 cells were transfected with pCIS-CK negative control vector or IFNγ driven pGAS reporter plasmid. 12 hrs after transfection, cells were either left untreated or treated with mBCD (5 mM, 30 min, 37°C) or infected with LD. Treated cells were then replenished or not with CH-Lipo. Subsequently, mBCD treated cells with or without CH-Lipo reloading were stimulated with or without 100U/ml of rIFNγ eight hours before final harvesting for luciferase activity determination in cell lysates. While the infected cells with or without CH-Lipo treatment were stimulated or not with 100 U/ml of rIFNγ at indicated timepoints post infection. Eight hours later, luciferase activities in cell lysates were measured as described in . The data represents the relative luciferase activity normalized to per microgram of protein content. Data is representative of three independent experiments.

Article Snippet: RAW 264.7 cells were transfected with the pGAS cis-reporter plasmids or the pCIS-CK negative control plasmid by electroporation by Genepulser X cell (Bio-Rad, Hercules, CA).

Techniques: Infection, Concentration Assay, Reporter Assay, Transfection, Negative Control, Plasmid Preparation, Luciferase, Activity Assay

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1) Product Images from "Restoration of IFNγR Subunit Assembly, IFNγ Signaling and Parasite Clearance in Leishmania donovani Infected Macrophages: Role of Membrane Cholesterol"

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