Structured Review

Toronto Research Chemicals 5 aminoimidazole 4 carboxamide riboside aicar
Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), <t>5-aminoimidazole-4-carboxamide</t> ribonucleotide <t>(AICAR),</t> 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.
5 Aminoimidazole 4 Carboxamide Riboside Aicar, supplied by Toronto Research Chemicals, used in various techniques. Bioz Stars score: 92/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Mass spectrometric analysis of purine de novo biosynthesis intermediates"

Article Title: Mass spectrometric analysis of purine de novo biosynthesis intermediates

Journal: PLoS ONE

doi: 10.1371/journal.pone.0208947

Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.
Figure Legend Snippet: Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.

Techniques Used:

2) Product Images from "AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression"

Article Title: AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression

Journal: Cell Death and Differentiation

doi: 10.1038/cdd.2017.14

CDX2 knockout abolishes the positive effects of AICAR on intestinal differentiation. Caco-2 cells were transfected with scrambled CRISPR/Cas9 plasmid (scramble-sgRNA) or CDX2 CRISPR/Cas9 plasmid (Cdx2 sgRNA) to delete Cdx2 or not. Transfected cells with GFP expression (plasmid carries GFP gene) were isolated using cell sorting, then treated with 0 or 0.2 mM AICAR for 4 days. ( a ) Protein contents of phospho-ACC, ACC, phospho-AMPK and AMPK. ( b ) Protein contents of CDX2, villin, E-cadherin and beta-actin. ( c ) Alkaline phosphatase activity. Data are representative of three separate experiments. Mean±S.E.M., n =3, * P
Figure Legend Snippet: CDX2 knockout abolishes the positive effects of AICAR on intestinal differentiation. Caco-2 cells were transfected with scrambled CRISPR/Cas9 plasmid (scramble-sgRNA) or CDX2 CRISPR/Cas9 plasmid (Cdx2 sgRNA) to delete Cdx2 or not. Transfected cells with GFP expression (plasmid carries GFP gene) were isolated using cell sorting, then treated with 0 or 0.2 mM AICAR for 4 days. ( a ) Protein contents of phospho-ACC, ACC, phospho-AMPK and AMPK. ( b ) Protein contents of CDX2, villin, E-cadherin and beta-actin. ( c ) Alkaline phosphatase activity. Data are representative of three separate experiments. Mean±S.E.M., n =3, * P

Techniques Used: Knock-Out, Transfection, CRISPR, Plasmid Preparation, Expressing, Isolation, FACS, Activity Assay

AMPK regulates Cdx2 transcription through inducing histone modifications. ( a ) Data analysis for epigenetic modifications in the Cdx2 promoter based on NCBI ChIP-Seq profile of H3K4me3 and H3K27me3. ( b ) CpG sites profile and genomic structure in the Cdx2 promoter. Blue regions show CpG islands. Red lines show CpG dinucleotides. −391 bp to −121 bp represents primer amplification region. ( c ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of AMPK VilCre KO and WT mice using ChIP-PCR. ( d ) H3K4me3 and H3K27me3 modifications in the Cdx2 promoter of Caco-2 cells treated with or without 0.2 mM AICAR and measured by ChIP-PCR. ( e ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of Caco-2 cells transfected with EGFP (CON), AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. ( f ) A proposed model for the effects of AMPK on intestinal differentiation via inducing histone modifications by regulating methylase and demethylase in the Cdx2 promoter. Data are representative of three separate experiments. Mean±S.E.M.; n =3, # P
Figure Legend Snippet: AMPK regulates Cdx2 transcription through inducing histone modifications. ( a ) Data analysis for epigenetic modifications in the Cdx2 promoter based on NCBI ChIP-Seq profile of H3K4me3 and H3K27me3. ( b ) CpG sites profile and genomic structure in the Cdx2 promoter. Blue regions show CpG islands. Red lines show CpG dinucleotides. −391 bp to −121 bp represents primer amplification region. ( c ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of AMPK VilCre KO and WT mice using ChIP-PCR. ( d ) H3K4me3 and H3K27me3 modifications in the Cdx2 promoter of Caco-2 cells treated with or without 0.2 mM AICAR and measured by ChIP-PCR. ( e ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of Caco-2 cells transfected with EGFP (CON), AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. ( f ) A proposed model for the effects of AMPK on intestinal differentiation via inducing histone modifications by regulating methylase and demethylase in the Cdx2 promoter. Data are representative of three separate experiments. Mean±S.E.M.; n =3, # P

Techniques Used: Chromatin Immunoprecipitation, Amplification, Binding Assay, Mouse Assay, Polymerase Chain Reaction, Transfection, Plasmid Preparation

AMPK enhances CDX2 expression in intestinal epithelial cells. Caco-2 cells were treated with 0 or 0.2 mM AICAR for 96 h when cells were collected for analysis. ( a ) Protein content of CDX2. ( b ) mRNA expression of Cdx2. ( c and d ) Immunofluorescent staining of CDX2 in non-transfected (NT) Caco-2 cells, or Caco-2 cells transfected with EGFP (CON), or AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. Scale bar is 100 μ m. Data are representative of three separate experiments. Mean±S.E.M., n =4, # P
Figure Legend Snippet: AMPK enhances CDX2 expression in intestinal epithelial cells. Caco-2 cells were treated with 0 or 0.2 mM AICAR for 96 h when cells were collected for analysis. ( a ) Protein content of CDX2. ( b ) mRNA expression of Cdx2. ( c and d ) Immunofluorescent staining of CDX2 in non-transfected (NT) Caco-2 cells, or Caco-2 cells transfected with EGFP (CON), or AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. Scale bar is 100 μ m. Data are representative of three separate experiments. Mean±S.E.M., n =4, # P

Techniques Used: Expressing, Staining, Transfection, Plasmid Preparation

AICAR treatment enhances differentiation of Caco-2 cells. ( a ) Alkaline phosphatase activity in Caco-2 cells treated with 0, 0.2 or 0.5 mM AICAR. ( b ) Enhanced protein content of phospho-AMPK, SI (sucrase-isomaltase), E-cadherin, DPPIV (dipeptidyl peptidase-4) and villin in Caco-2 cells treated with 0.2 or 0.5 mM AICAR. ( c ) Transepithelial electrical resistance over time. ( d ) FITC-dextran paracellular intestinal epithelial permeability at 21 days post incubation. ( e ) Immunofluorescence staining of tight junction protein ZO-1 pre- and post- calcium switch assay. Caco-2 cells were grown to confluence with or without 0.2 mM AICAR and subjected to a calcium switch assay. Cells were fixed at various time points (0, 8, 16 and 24 h) after restoration of Ca 2+ containing medium. Data are representative of three separate experiments. Scale bar is 100 μ m. Mean±S.E.M., n =4, * P
Figure Legend Snippet: AICAR treatment enhances differentiation of Caco-2 cells. ( a ) Alkaline phosphatase activity in Caco-2 cells treated with 0, 0.2 or 0.5 mM AICAR. ( b ) Enhanced protein content of phospho-AMPK, SI (sucrase-isomaltase), E-cadherin, DPPIV (dipeptidyl peptidase-4) and villin in Caco-2 cells treated with 0.2 or 0.5 mM AICAR. ( c ) Transepithelial electrical resistance over time. ( d ) FITC-dextran paracellular intestinal epithelial permeability at 21 days post incubation. ( e ) Immunofluorescence staining of tight junction protein ZO-1 pre- and post- calcium switch assay. Caco-2 cells were grown to confluence with or without 0.2 mM AICAR and subjected to a calcium switch assay. Cells were fixed at various time points (0, 8, 16 and 24 h) after restoration of Ca 2+ containing medium. Data are representative of three separate experiments. Scale bar is 100 μ m. Mean±S.E.M., n =4, * P

Techniques Used: Activity Assay, Permeability, Incubation, Immunofluorescence, Staining

3) Product Images from "Mass spectrometric analysis of purine de novo biosynthesis intermediates"

Article Title: Mass spectrometric analysis of purine de novo biosynthesis intermediates

Journal: PLoS ONE

doi: 10.1371/journal.pone.0208947

Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.
Figure Legend Snippet: Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.

Techniques Used:

4) Product Images from "AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression"

Article Title: AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression

Journal: Cell Death and Differentiation

doi: 10.1038/cdd.2017.14

CDX2 knockout abolishes the positive effects of AICAR on intestinal differentiation. Caco-2 cells were transfected with scrambled CRISPR/Cas9 plasmid (scramble-sgRNA) or CDX2 CRISPR/Cas9 plasmid (Cdx2 sgRNA) to delete Cdx2 or not. Transfected cells with GFP expression (plasmid carries GFP gene) were isolated using cell sorting, then treated with 0 or 0.2 mM AICAR for 4 days. ( a ) Protein contents of phospho-ACC, ACC, phospho-AMPK and AMPK. ( b ) Protein contents of CDX2, villin, E-cadherin and beta-actin. ( c ) Alkaline phosphatase activity. Data are representative of three separate experiments. Mean±S.E.M., n =3, * P
Figure Legend Snippet: CDX2 knockout abolishes the positive effects of AICAR on intestinal differentiation. Caco-2 cells were transfected with scrambled CRISPR/Cas9 plasmid (scramble-sgRNA) or CDX2 CRISPR/Cas9 plasmid (Cdx2 sgRNA) to delete Cdx2 or not. Transfected cells with GFP expression (plasmid carries GFP gene) were isolated using cell sorting, then treated with 0 or 0.2 mM AICAR for 4 days. ( a ) Protein contents of phospho-ACC, ACC, phospho-AMPK and AMPK. ( b ) Protein contents of CDX2, villin, E-cadherin and beta-actin. ( c ) Alkaline phosphatase activity. Data are representative of three separate experiments. Mean±S.E.M., n =3, * P

Techniques Used: Knock-Out, Transfection, CRISPR, Plasmid Preparation, Expressing, Isolation, FACS, Activity Assay

AMPK regulates Cdx2 transcription through inducing histone modifications. ( a ) Data analysis for epigenetic modifications in the Cdx2 promoter based on NCBI ChIP-Seq profile of H3K4me3 and H3K27me3. ( b ) CpG sites profile and genomic structure in the Cdx2 promoter. Blue regions show CpG islands. Red lines show CpG dinucleotides. −391 bp to −121 bp represents primer amplification region. ( c ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of AMPK VilCre KO and WT mice using ChIP-PCR. ( d ) H3K4me3 and H3K27me3 modifications in the Cdx2 promoter of Caco-2 cells treated with or without 0.2 mM AICAR and measured by ChIP-PCR. ( e ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of Caco-2 cells transfected with EGFP (CON), AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. ( f ) A proposed model for the effects of AMPK on intestinal differentiation via inducing histone modifications by regulating methylase and demethylase in the Cdx2 promoter. Data are representative of three separate experiments. Mean±S.E.M.; n =3, # P
Figure Legend Snippet: AMPK regulates Cdx2 transcription through inducing histone modifications. ( a ) Data analysis for epigenetic modifications in the Cdx2 promoter based on NCBI ChIP-Seq profile of H3K4me3 and H3K27me3. ( b ) CpG sites profile and genomic structure in the Cdx2 promoter. Blue regions show CpG islands. Red lines show CpG dinucleotides. −391 bp to −121 bp represents primer amplification region. ( c ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of AMPK VilCre KO and WT mice using ChIP-PCR. ( d ) H3K4me3 and H3K27me3 modifications in the Cdx2 promoter of Caco-2 cells treated with or without 0.2 mM AICAR and measured by ChIP-PCR. ( e ) H3K4me3 and H3K27me3 modifications as well as binding status of EZH2 and LSD1 in the Cdx2 promoter of Caco-2 cells transfected with EGFP (CON), AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. ( f ) A proposed model for the effects of AMPK on intestinal differentiation via inducing histone modifications by regulating methylase and demethylase in the Cdx2 promoter. Data are representative of three separate experiments. Mean±S.E.M.; n =3, # P

Techniques Used: Chromatin Immunoprecipitation, Amplification, Binding Assay, Mouse Assay, Polymerase Chain Reaction, Transfection, Plasmid Preparation

AMPK promotes intestinal epithelial differentiation. Caco-2 cells were transfected with EGFP (CON), AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. ( a ) Phospho-ACC and phospho-AMPK activation. ( b ) Transepithelial electrical resistance over time. ( c ) FITC-dextran paracellular intestinal epithelial permeability at 21 days post incubation. ( d ) Immunofluorescent staining of tight junction protein ZO-1 pre- and post- calcium switch assay. Caco-2 cells were grown to confluence and subjected to a calcium switch assay. Cells were fixed 18 h before calcium switch (−18 h) and various time points (0, 4, 8, 16 and 24 h) after restoration of Ca 2+ containing medium. Scale bar is 100 μ m. ( e ) Alkaline phosphatase activity. ( f ) Protein contents of SI (sucrase-isomaltase), E-cadherin, DPPIV (dipeptidyl peptidase-4) and villin. Data are representative of three separate experiments. Mean±S.E.M., n =4, # P
Figure Legend Snippet: AMPK promotes intestinal epithelial differentiation. Caco-2 cells were transfected with EGFP (CON), AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. ( a ) Phospho-ACC and phospho-AMPK activation. ( b ) Transepithelial electrical resistance over time. ( c ) FITC-dextran paracellular intestinal epithelial permeability at 21 days post incubation. ( d ) Immunofluorescent staining of tight junction protein ZO-1 pre- and post- calcium switch assay. Caco-2 cells were grown to confluence and subjected to a calcium switch assay. Cells were fixed 18 h before calcium switch (−18 h) and various time points (0, 4, 8, 16 and 24 h) after restoration of Ca 2+ containing medium. Scale bar is 100 μ m. ( e ) Alkaline phosphatase activity. ( f ) Protein contents of SI (sucrase-isomaltase), E-cadherin, DPPIV (dipeptidyl peptidase-4) and villin. Data are representative of three separate experiments. Mean±S.E.M., n =4, # P

Techniques Used: Transfection, Plasmid Preparation, Activation Assay, Permeability, Incubation, Staining, Activity Assay

AMPK enhances CDX2 expression in intestinal epithelial cells. Caco-2 cells were treated with 0 or 0.2 mM AICAR for 96 h when cells were collected for analysis. ( a ) Protein content of CDX2. ( b ) mRNA expression of Cdx2. ( c and d ) Immunofluorescent staining of CDX2 in non-transfected (NT) Caco-2 cells, or Caco-2 cells transfected with EGFP (CON), or AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. Scale bar is 100 μ m. Data are representative of three separate experiments. Mean±S.E.M., n =4, # P
Figure Legend Snippet: AMPK enhances CDX2 expression in intestinal epithelial cells. Caco-2 cells were treated with 0 or 0.2 mM AICAR for 96 h when cells were collected for analysis. ( a ) Protein content of CDX2. ( b ) mRNA expression of Cdx2. ( c and d ) Immunofluorescent staining of CDX2 in non-transfected (NT) Caco-2 cells, or Caco-2 cells transfected with EGFP (CON), or AMPK α WT (WT) or AMPK α K45R (K45R) plasmid. Scale bar is 100 μ m. Data are representative of three separate experiments. Mean±S.E.M., n =4, # P

Techniques Used: Expressing, Staining, Transfection, Plasmid Preparation

AICAR treatment enhances differentiation of Caco-2 cells. ( a ) Alkaline phosphatase activity in Caco-2 cells treated with 0, 0.2 or 0.5 mM AICAR. ( b ) Enhanced protein content of phospho-AMPK, SI (sucrase-isomaltase), E-cadherin, DPPIV (dipeptidyl peptidase-4) and villin in Caco-2 cells treated with 0.2 or 0.5 mM AICAR. ( c ) Transepithelial electrical resistance over time. ( d ) FITC-dextran paracellular intestinal epithelial permeability at 21 days post incubation. ( e ) Immunofluorescence staining of tight junction protein ZO-1 pre- and post- calcium switch assay. Caco-2 cells were grown to confluence with or without 0.2 mM AICAR and subjected to a calcium switch assay. Cells were fixed at various time points (0, 8, 16 and 24 h) after restoration of Ca 2+ containing medium. Data are representative of three separate experiments. Scale bar is 100 μ m. Mean±S.E.M., n =4, * P
Figure Legend Snippet: AICAR treatment enhances differentiation of Caco-2 cells. ( a ) Alkaline phosphatase activity in Caco-2 cells treated with 0, 0.2 or 0.5 mM AICAR. ( b ) Enhanced protein content of phospho-AMPK, SI (sucrase-isomaltase), E-cadherin, DPPIV (dipeptidyl peptidase-4) and villin in Caco-2 cells treated with 0.2 or 0.5 mM AICAR. ( c ) Transepithelial electrical resistance over time. ( d ) FITC-dextran paracellular intestinal epithelial permeability at 21 days post incubation. ( e ) Immunofluorescence staining of tight junction protein ZO-1 pre- and post- calcium switch assay. Caco-2 cells were grown to confluence with or without 0.2 mM AICAR and subjected to a calcium switch assay. Cells were fixed at various time points (0, 8, 16 and 24 h) after restoration of Ca 2+ containing medium. Data are representative of three separate experiments. Scale bar is 100 μ m. Mean±S.E.M., n =4, * P

Techniques Used: Activity Assay, Permeability, Incubation, Immunofluorescence, Staining

Related Articles

In Vitro:

Article Title: AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression
Article Snippet: .. Unless specified, the medium was refreshed every 48 h. For AICAR treatment, Caco-2 cells were seeded at a density of 2 × 105 per well on 12-well plates, and treated with 0, 0.2 or 0.5 mM of 5-aminoimidazole-4-carboxamide riboside (AICAR; Toronto Research Chemical Inc., Ontario, Canada) for 0, 12, 24, 48, and 96 h (for immunoblotting assay and mRNA assay) as well as 4 days (for alkaline phosphatase assay, immunoblotting assay and immunofluorescent staining), 7 days (for alkaline phosphatase assay) and 21 days (for in vitro barrier function assessment). .. For plasmid transfection, Caco-2 cells were transfected with plasmids using Lipofectamine 3000 (Life Technologies) per manufacturer's instructions.

ALP Assay:

Article Title: AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression
Article Snippet: .. Unless specified, the medium was refreshed every 48 h. For AICAR treatment, Caco-2 cells were seeded at a density of 2 × 105 per well on 12-well plates, and treated with 0, 0.2 or 0.5 mM of 5-aminoimidazole-4-carboxamide riboside (AICAR; Toronto Research Chemical Inc., Ontario, Canada) for 0, 12, 24, 48, and 96 h (for immunoblotting assay and mRNA assay) as well as 4 days (for alkaline phosphatase assay, immunoblotting assay and immunofluorescent staining), 7 days (for alkaline phosphatase assay) and 21 days (for in vitro barrier function assessment). .. For plasmid transfection, Caco-2 cells were transfected with plasmids using Lipofectamine 3000 (Life Technologies) per manufacturer's instructions.

Incubation:

Article Title: Regulation of skeletal muscle sucrose, non-fermenting 1/AMP-activated protein kinase-related kinase (SNARK) by metabolic stress and diabetes
Article Snippet: .. Cells were washed once with PBS and incubated for 5.5 h with 1 ml DMEM (1 g glucose/l), supplemented with fatty acid-free BSA (0.2% wt/vol.) and [9,10(n )-3 H]palmitic acid (1850 Bq/ml; Amersham), with or without insulin (120 nmol/l) or AICAR (1 mmol/l; Toronto Research Chemicals, Toronto, ON, Canada). .. To absorb non-metabolised palmitate, 0.2 ml of the cell supernatant was mixed with 0.8 ml charcoal slurry (0.1 g charcoal powder in 1 ml 0.02 mol/l TRIS–HCl buffer, pH 7.5) in a 2 ml Eppendorf tube and shaken for 30 min.

other:

Article Title: 5′AMP-activated protein kinase α deficiency enhances stress-induced apoptosis in BHK and PC12 cells
Article Snippet: The AMPK activator, AICAR, was obtained from Toronto Research Chemicals, solubilized in water and used at working concentrations of 100 μM and 1 mM.

Expressing:

Article Title: Autophagy Stimulus Promotes Early HuR Protein Activation and p62/SQSTM1 Protein Synthesis in ARPE-19 Cells by Triggering Erk1/2, p38MAPK, and JNK Kinase Pathways
Article Snippet: .. To find out the best conditions for studying both HuR protein translocation and p62 protein expression, cells were exposed to either the solvent (DMSO, 0.1%), the proteasome inhibitor MG132 (1 μ M, Calbiochem, San Diego, CA), or AICAR (2 mM 5-aminoimidazole-4-carboxy amide ribonucleoside, Toronto Research Chemical, Canada), alone or together, for 15 min, 30 min, or 2 hrs. ..

Western Blot:

Article Title: 5-aminoimidazole-4-carboxamide Riboside Induces Apoptosis Through AMP-activated Protein Kinase-independent and NADPH Oxidase-dependent Pathways
Article Snippet: .. Western blot analysis Jurkat and THP1 cells were treated with or without 5 mM AICAR (Toronto Research Chemicals, Inc) for different times, as indicated in each experiment. .. Cells were washed once with PBS and lysed in sample buffer (80~120 µl), and then immediately boiled for 5 min. Each of the samples was resolved in 10 or 12% SDS-PAGE, after which the proteins were electrotransferred onto a polyvinylidene difluoride (PVDF) membrane (GE Healthcare, Waukesha, WI).

Staining:

Article Title: AMPK improves gut epithelial differentiation and barrier function via regulating Cdx2 expression
Article Snippet: .. Unless specified, the medium was refreshed every 48 h. For AICAR treatment, Caco-2 cells were seeded at a density of 2 × 105 per well on 12-well plates, and treated with 0, 0.2 or 0.5 mM of 5-aminoimidazole-4-carboxamide riboside (AICAR; Toronto Research Chemical Inc., Ontario, Canada) for 0, 12, 24, 48, and 96 h (for immunoblotting assay and mRNA assay) as well as 4 days (for alkaline phosphatase assay, immunoblotting assay and immunofluorescent staining), 7 days (for alkaline phosphatase assay) and 21 days (for in vitro barrier function assessment). .. For plasmid transfection, Caco-2 cells were transfected with plasmids using Lipofectamine 3000 (Life Technologies) per manufacturer's instructions.

Translocation Assay:

Article Title: Autophagy Stimulus Promotes Early HuR Protein Activation and p62/SQSTM1 Protein Synthesis in ARPE-19 Cells by Triggering Erk1/2, p38MAPK, and JNK Kinase Pathways
Article Snippet: .. To find out the best conditions for studying both HuR protein translocation and p62 protein expression, cells were exposed to either the solvent (DMSO, 0.1%), the proteasome inhibitor MG132 (1 μ M, Calbiochem, San Diego, CA), or AICAR (2 mM 5-aminoimidazole-4-carboxy amide ribonucleoside, Toronto Research Chemical, Canada), alone or together, for 15 min, 30 min, or 2 hrs. ..

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    Toronto Research Chemicals aica riboside
    The combined effect of MTX and <t>AICA</t> <t>riboside</t> on inhibition of cancer cell DNA synthesis. Data are presented as mean±SD ( n =3). c P
    Aica Riboside, supplied by Toronto Research Chemicals, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aica riboside/product/Toronto Research Chemicals
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    Toronto Research Chemicals 5 aminoimidazole 4 carboxamide riboside aicar
    Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), <t>5-aminoimidazole-4-carboxamide</t> ribonucleotide <t>(AICAR),</t> 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.
    5 Aminoimidazole 4 Carboxamide Riboside Aicar, supplied by Toronto Research Chemicals, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5 aminoimidazole 4 carboxamide riboside aicar/product/Toronto Research Chemicals
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
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    92
    Toronto Research Chemicals aicar
    Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), <t>5-aminoimidazole-4-carboxamide</t> ribonucleotide <t>(AICAR),</t> 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.
    Aicar, supplied by Toronto Research Chemicals, used in various techniques. Bioz Stars score: 92/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The combined effect of MTX and AICA riboside on inhibition of cancer cell DNA synthesis. Data are presented as mean±SD ( n =3). c P

    Journal: Acta Pharmacologica Sinica

    Article Title: Methotrexate and 5-aminoimidazole-4-carboxamide riboside exert synergistic anticancer action against human breast cancer and hepatocellular carcinoma

    doi: 10.1038/aps.2013.16

    Figure Lengend Snippet: The combined effect of MTX and AICA riboside on inhibition of cancer cell DNA synthesis. Data are presented as mean±SD ( n =3). c P

    Article Snippet: AICA riboside and AICA ribotide were obtained from Toronto Research Chemicals (Toronto, Canada).

    Techniques: Inhibition, DNA Synthesis

    Effect of MTX, AICA riboside and their combination on the growth of MCF-7 cell xenografts in BALB/c nude mice. Data are presented as mean±SD ( n =8). b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Methotrexate and 5-aminoimidazole-4-carboxamide riboside exert synergistic anticancer action against human breast cancer and hepatocellular carcinoma

    doi: 10.1038/aps.2013.16

    Figure Lengend Snippet: Effect of MTX, AICA riboside and their combination on the growth of MCF-7 cell xenografts in BALB/c nude mice. Data are presented as mean±SD ( n =8). b P

    Article Snippet: AICA riboside and AICA ribotide were obtained from Toronto Research Chemicals (Toronto, Canada).

    Techniques: Mouse Assay

    Synergistic effect of MTX and AICA riboside on inhibition of MCF-7 (A) and HepG2 (B) cell proliferation in vitro . CI

    Journal: Acta Pharmacologica Sinica

    Article Title: Methotrexate and 5-aminoimidazole-4-carboxamide riboside exert synergistic anticancer action against human breast cancer and hepatocellular carcinoma

    doi: 10.1038/aps.2013.16

    Figure Lengend Snippet: Synergistic effect of MTX and AICA riboside on inhibition of MCF-7 (A) and HepG2 (B) cell proliferation in vitro . CI

    Article Snippet: AICA riboside and AICA ribotide were obtained from Toronto Research Chemicals (Toronto, Canada).

    Techniques: Inhibition, In Vitro

    Effect of MTX, AICA riboside, and their combination on AMPK phosphorylation and activity. (A) 100 μg protein was subjected to Western blot analysis with antibody against phospho-Thr172 AMPK α (pAMPK) or β-actin. Results are representative of three independent experiments. (B) Purified proteins were assayed for their ability to phosphorylate SAMS in vitro in the presence of [γ- 32 P]ATP. Values were corrected for differences in protein concentration, and expressed as mean±SD ( n =2). b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Methotrexate and 5-aminoimidazole-4-carboxamide riboside exert synergistic anticancer action against human breast cancer and hepatocellular carcinoma

    doi: 10.1038/aps.2013.16

    Figure Lengend Snippet: Effect of MTX, AICA riboside, and their combination on AMPK phosphorylation and activity. (A) 100 μg protein was subjected to Western blot analysis with antibody against phospho-Thr172 AMPK α (pAMPK) or β-actin. Results are representative of three independent experiments. (B) Purified proteins were assayed for their ability to phosphorylate SAMS in vitro in the presence of [γ- 32 P]ATP. Values were corrected for differences in protein concentration, and expressed as mean±SD ( n =2). b P

    Article Snippet: AICA riboside and AICA ribotide were obtained from Toronto Research Chemicals (Toronto, Canada).

    Techniques: Activity Assay, Western Blot, Purification, In Vitro, Protein Concentration

    Combined effects of MTX and AICA riboside on inhibition of MCF-7 (A) and HepG2 (B) cell proliferation in vitro . Data are presented as mean±SD ( n =6). c P

    Journal: Acta Pharmacologica Sinica

    Article Title: Methotrexate and 5-aminoimidazole-4-carboxamide riboside exert synergistic anticancer action against human breast cancer and hepatocellular carcinoma

    doi: 10.1038/aps.2013.16

    Figure Lengend Snippet: Combined effects of MTX and AICA riboside on inhibition of MCF-7 (A) and HepG2 (B) cell proliferation in vitro . Data are presented as mean±SD ( n =6). c P

    Article Snippet: AICA riboside and AICA ribotide were obtained from Toronto Research Chemicals (Toronto, Canada).

    Techniques: Inhibition, In Vitro

    Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.

    Journal: PLoS ONE

    Article Title: Mass spectrometric analysis of purine de novo biosynthesis intermediates

    doi: 10.1371/journal.pone.0208947

    Figure Lengend Snippet: Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.

    Article Snippet: AICAR, 5-aminoimidazole-4-carboxamide riboside (AICAr) and adenylosuccinic acid (SAMP) were purchased from Toronto Research Chemicals Inc. (North York, Canada).

    Techniques:

    Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.

    Journal: PLoS ONE

    Article Title: Mass spectrometric analysis of purine de novo biosynthesis intermediates

    doi: 10.1371/journal.pone.0208947

    Figure Lengend Snippet: Purine de novo synthesis in humans. Abbreviations: phosphoribosylamine (PRA), glycinamideribonucleotide (GAR), N-formylglycinamide ribonucleotide (FGAR), N-formylglycinamidine ribonucleotide (FGAMR), aminoimidazole ribonucleotide (AIR), carboxyaminoimidazole ribonucleotide (CAIR), N-succinocarboxamide-5-aminoimidazole ribonucleotide (SAICAR), 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR), inosine-5-monophosphate (IMP), glutamine (Gln), glutamate (Glu), pyrophosphate (PPi), adenosine-5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), glycine (Gly), phosphate (Pi), N 10 -formyl tetrahydrofolate (N 10 -formyl THF), tetrahydrofolate (THF), hydrogen carbonic acid (HCO 3- ), aspartate (Asp). For enzyme abbreviations, see the second paragraph of the Introduction.

    Article Snippet: AICAR, 5-aminoimidazole-4-carboxamide riboside (AICAr) and adenylosuccinic acid (SAMP) were purchased from Toronto Research Chemicals Inc. (North York, Canada).

    Techniques: