rabbit igg anti idh1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit igg anti idh1
    <t>IDH1</t> catalyzes the conversion of isocitrate to αKG.
    Rabbit Igg Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit igg anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit igg anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1"

    Article Title: A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1

    Journal: The Analyst

    doi: 10.1039/d0an00174k

    IDH1 catalyzes the conversion of isocitrate to αKG.
    Figure Legend Snippet: IDH1 catalyzes the conversion of isocitrate to αKG.

    Techniques Used:

    (a) Scheme for the preparation and analysis of recombinant IDH1 samples. (b) αKG concentrations measured in the 384-sample biochemical screen (7 samples did not produce either 12C or 13C αKG capture).
    Figure Legend Snippet: (a) Scheme for the preparation and analysis of recombinant IDH1 samples. (b) αKG concentrations measured in the 384-sample biochemical screen (7 samples did not produce either 12C or 13C αKG capture).

    Techniques Used: Recombinant

    IDH1 expression level as determined with western blot and αKG concentration as measured with SAMDI-MS for untreated cells and cells treated with transfection agent, control sequence siRNA and IDH1-targeting siRNA. All data are expressed as mean ± standard deviation with n = 2 experimental replicates.
    Figure Legend Snippet: IDH1 expression level as determined with western blot and αKG concentration as measured with SAMDI-MS for untreated cells and cells treated with transfection agent, control sequence siRNA and IDH1-targeting siRNA. All data are expressed as mean ± standard deviation with n = 2 experimental replicates.

    Techniques Used: Expressing, Western Blot, Concentration Assay, Transfection, Sequencing, Standard Deviation

    rabbit igg anti idh1  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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  • 93

    Structured Review

    Cell Signaling Technology Inc rabbit igg anti idh1
    <t>IDH1</t> catalyzes the conversion of isocitrate to αKG.
    Rabbit Igg Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit igg anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit igg anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1"

    Article Title: A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1

    Journal: The Analyst

    doi: 10.1039/d0an00174k

    IDH1 catalyzes the conversion of isocitrate to αKG.
    Figure Legend Snippet: IDH1 catalyzes the conversion of isocitrate to αKG.

    Techniques Used:

    (a) Scheme for the preparation and analysis of recombinant IDH1 samples. (b) αKG concentrations measured in the 384-sample biochemical screen (7 samples did not produce either 12C or 13C αKG capture).
    Figure Legend Snippet: (a) Scheme for the preparation and analysis of recombinant IDH1 samples. (b) αKG concentrations measured in the 384-sample biochemical screen (7 samples did not produce either 12C or 13C αKG capture).

    Techniques Used: Recombinant

    IDH1 expression level as determined with western blot and αKG concentration as measured with SAMDI-MS for untreated cells and cells treated with transfection agent, control sequence siRNA and IDH1-targeting siRNA. All data are expressed as mean ± standard deviation with n = 2 experimental replicates.
    Figure Legend Snippet: IDH1 expression level as determined with western blot and αKG concentration as measured with SAMDI-MS for untreated cells and cells treated with transfection agent, control sequence siRNA and IDH1-targeting siRNA. All data are expressed as mean ± standard deviation with n = 2 experimental replicates.

    Techniques Used: Expressing, Western Blot, Concentration Assay, Transfection, Sequencing, Standard Deviation

    rabbit anti idh1  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Structured Review

    Cell Signaling Technology Inc rabbit anti idh1
    Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting <t>IDH1</t> or treated with 15μM GSK864 (IDH1i). Empty vector or DMSO was used as a control, respectively.
    Rabbit Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer"

    Article Title: Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer

    Journal: Molecular cancer research : MCR

    doi: 10.1158/1541-7786.MCR-18-1233

    Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting IDH1 or treated with 15μM GSK864 (IDH1i). Empty vector or DMSO was used as a control, respectively.
    Figure Legend Snippet: Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting IDH1 or treated with 15μM GSK864 (IDH1i). Empty vector or DMSO was used as a control, respectively.

    Techniques Used: Infection, Expressing, Plasmid Preparation

    (A-F) Ovcar3 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control.
    Figure Legend Snippet: (A-F) Ovcar3 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control.

    Techniques Used: Infection, Expressing, Plasmid Preparation

    Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Empty vector was used as a control.
    Figure Legend Snippet: Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Empty vector was used as a control.

    Techniques Used: Infection, Plasmid Preparation

    rabbit igg anti idh1  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
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  • 93

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    Cell Signaling Technology Inc rabbit igg anti idh1
    Rabbit Igg Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit igg anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit igg anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

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    monoclonal antibodies against idh1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc monoclonal antibodies against idh1
    Genetic mutations in <t> isocitrate dehydrogenase </t> (IDH)1/2
    Monoclonal Antibodies Against Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal antibodies against idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal antibodies against idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Pharmacological characterization of TQ 05310, a potent inhibitor of isocitrate dehydrogenase 2 R140Q and R172K mutants"

    Article Title: Pharmacological characterization of TQ 05310, a potent inhibitor of isocitrate dehydrogenase 2 R140Q and R172K mutants

    Journal: Cancer Science

    doi: 10.1111/cas.14152

    Genetic mutations in  isocitrate dehydrogenase  (IDH)1/2
    Figure Legend Snippet: Genetic mutations in isocitrate dehydrogenase (IDH)1/2

    Techniques Used: Mutagenesis

    anti idh1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti idh1

    Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Polyamines and eIF5A Hypusination Modulate Mitochondrial Respiration and Macrophage Activation"

    Article Title: Polyamines and eIF5A Hypusination Modulate Mitochondrial Respiration and Macrophage Activation

    Journal: Cell Metabolism

    doi: 10.1016/j.cmet.2019.05.003


    Figure Legend Snippet:

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, Construct, Plasmid Preparation, Sequencing, Clone Assay, Software

    phycoerythrin pe conjugated anti idh1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phycoerythrin pe conjugated anti idh1
    Proteome-transcriptome correlation in human hematopoietic stem and progenitor cell subpopulations. ( A ) Nonsupervised hierarchical clustering (Euclidean distance) heatmap of the intensity for the transcripts identified in HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs (shades of red) isolated from five different HSPC donors (shades of blue). The transcript intensities are centered and scaled and depicted in color shades from red to blue. The transcripts with missing transcript intensity values in all samples were removed because they could not be handled by the clustering algorithm. Remaining missing transcript intensity values are shown in white. Clustering was observed mostly according to cell type, not according to donor. ( B ) GO enrichment analysis showed good alignment of protein and mRNA data. GSEA was performed for ranked mRNA and protein lists using GO processes from the Gene Ontology Consortium database as gene sets. Shown are normalized enrichment scores for the individual gene sets. Significantly up-regulated gene sets are marked in blue color; significantly downregulated gene sets are marked in red color. Significance was defined as FDR < 0.25, specific cell subpopulations were compared with the average of the remaining three cell types, and log2(fold change) was used as ranking criterion. Empty fields mean that no enrichment could be calculated. Abbreviations: MEGA, megakaryocyte; MAPK, mitogen-activated protein kinase; PI3K, phosphoinositide-3-kinase; PLC, phospholipase C. ( C ) Correlation between proteomics and transcriptomics data for HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs. Dots are depicted in red when the FDR was below 0.01 both for protein and RNA data, orange when FDR < 0.01 for protein data, purple when FDR < 0.01 for RNA data, and gray when FDR ≥ 0.01 for both protein and RNA data. ( D ) Network analysis of significantly up-regulated proteins with concomitant significantly downregulated mRNA in HSCs/MPPs. Two clusters were especially prominent, including the snoRNPs and telomerase complex proteins GAR1, DKC1, NOP10, NHP2, and the quiescence-inducing NAD(P)H-producing proteins <t>IDH1,</t> IDH3A, and IDH3B. HSCs/MPPs were compared with the average of the other three subpopulations; cutoffs were set at FDR < 0.01 for protein and RNA data. Colors depict GO terms found enriched in B .
    Phycoerythrin Pe Conjugated Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phycoerythrin pe conjugated anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phycoerythrin pe conjugated anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Sensitive Quantitative Proteomics of Human Hematopoietic Stem and Progenitor Cells by Data-independent Acquisition Mass Spectrometry"

    Article Title: Sensitive Quantitative Proteomics of Human Hematopoietic Stem and Progenitor Cells by Data-independent Acquisition Mass Spectrometry

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1074/mcp.TIR119.001431

    Proteome-transcriptome correlation in human hematopoietic stem and progenitor cell subpopulations. ( A ) Nonsupervised hierarchical clustering (Euclidean distance) heatmap of the intensity for the transcripts identified in HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs (shades of red) isolated from five different HSPC donors (shades of blue). The transcript intensities are centered and scaled and depicted in color shades from red to blue. The transcripts with missing transcript intensity values in all samples were removed because they could not be handled by the clustering algorithm. Remaining missing transcript intensity values are shown in white. Clustering was observed mostly according to cell type, not according to donor. ( B ) GO enrichment analysis showed good alignment of protein and mRNA data. GSEA was performed for ranked mRNA and protein lists using GO processes from the Gene Ontology Consortium database as gene sets. Shown are normalized enrichment scores for the individual gene sets. Significantly up-regulated gene sets are marked in blue color; significantly downregulated gene sets are marked in red color. Significance was defined as FDR < 0.25, specific cell subpopulations were compared with the average of the remaining three cell types, and log2(fold change) was used as ranking criterion. Empty fields mean that no enrichment could be calculated. Abbreviations: MEGA, megakaryocyte; MAPK, mitogen-activated protein kinase; PI3K, phosphoinositide-3-kinase; PLC, phospholipase C. ( C ) Correlation between proteomics and transcriptomics data for HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs. Dots are depicted in red when the FDR was below 0.01 both for protein and RNA data, orange when FDR < 0.01 for protein data, purple when FDR < 0.01 for RNA data, and gray when FDR ≥ 0.01 for both protein and RNA data. ( D ) Network analysis of significantly up-regulated proteins with concomitant significantly downregulated mRNA in HSCs/MPPs. Two clusters were especially prominent, including the snoRNPs and telomerase complex proteins GAR1, DKC1, NOP10, NHP2, and the quiescence-inducing NAD(P)H-producing proteins IDH1, IDH3A, and IDH3B. HSCs/MPPs were compared with the average of the other three subpopulations; cutoffs were set at FDR < 0.01 for protein and RNA data. Colors depict GO terms found enriched in B .
    Figure Legend Snippet: Proteome-transcriptome correlation in human hematopoietic stem and progenitor cell subpopulations. ( A ) Nonsupervised hierarchical clustering (Euclidean distance) heatmap of the intensity for the transcripts identified in HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs (shades of red) isolated from five different HSPC donors (shades of blue). The transcript intensities are centered and scaled and depicted in color shades from red to blue. The transcripts with missing transcript intensity values in all samples were removed because they could not be handled by the clustering algorithm. Remaining missing transcript intensity values are shown in white. Clustering was observed mostly according to cell type, not according to donor. ( B ) GO enrichment analysis showed good alignment of protein and mRNA data. GSEA was performed for ranked mRNA and protein lists using GO processes from the Gene Ontology Consortium database as gene sets. Shown are normalized enrichment scores for the individual gene sets. Significantly up-regulated gene sets are marked in blue color; significantly downregulated gene sets are marked in red color. Significance was defined as FDR < 0.25, specific cell subpopulations were compared with the average of the remaining three cell types, and log2(fold change) was used as ranking criterion. Empty fields mean that no enrichment could be calculated. Abbreviations: MEGA, megakaryocyte; MAPK, mitogen-activated protein kinase; PI3K, phosphoinositide-3-kinase; PLC, phospholipase C. ( C ) Correlation between proteomics and transcriptomics data for HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs. Dots are depicted in red when the FDR was below 0.01 both for protein and RNA data, orange when FDR < 0.01 for protein data, purple when FDR < 0.01 for RNA data, and gray when FDR ≥ 0.01 for both protein and RNA data. ( D ) Network analysis of significantly up-regulated proteins with concomitant significantly downregulated mRNA in HSCs/MPPs. Two clusters were especially prominent, including the snoRNPs and telomerase complex proteins GAR1, DKC1, NOP10, NHP2, and the quiescence-inducing NAD(P)H-producing proteins IDH1, IDH3A, and IDH3B. HSCs/MPPs were compared with the average of the other three subpopulations; cutoffs were set at FDR < 0.01 for protein and RNA data. Colors depict GO terms found enriched in B .

    Techniques Used: Isolation

    rabbit anti idh1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti idh1
    (A) LC/MS comparison of 2 fallopian tube (FT282 and FT4-Tag) and 2 HGSC (Ovcar3 and Ovcar10) cell lines. Heat map shows z-scores of relative abundance normalized to the mean of each individual TCA cycle metabolite. Significant metabolites are labeled in green. Adjusted p-value (FDR) <0.05. (B) Differentially expressed transcripts in 5 parental fallopian tube (FT282, FT4-Tag, FT6-Tag, FT33-Tag, and PFTE4) and 4 HGSC (HeyA8, Ovcar5, Ovcar3, and Ovcar10) cell lines. Heat map shows z-scores of expression levels relative to B2M of 27 TCA cycle enzymes. Significant transcripts are labeled in green. Adjusted p-value (FDR) <0.05 (C) Differentially expressed transcripts of 27 TCA cycle enzymes. Heat map shows fold change z-scores of indicated HGSC cell lines cultured in ultra-low attachment and adherent conditions. Significant transcripts are labeled in green. Adjusted p-value (FDR) <0.001 (D) Kaplan-Meier curve for disease progression-free survival of ovarian cancer patients classified according to <t>IDH1</t> expression level. Ovarian cancer patients were filtered for a serous histosubtype and TP53 mutation. Estimates of the hazard ration (HR) and logrank p-values are indicated. (E) Immunoblot analysis of IDH1 comparing normal human fallopian tube and HGSC patient tissue lysates. Vinculin was used as a loading control. (F) IDH1 protein expression normalized to vinculin and quantified using ImageJ. Data represent mean ± SD. *p<0.03.
    Rabbit Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer"

    Article Title: Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer

    Journal: bioRxiv

    doi: 10.1101/472613

    (A) LC/MS comparison of 2 fallopian tube (FT282 and FT4-Tag) and 2 HGSC (Ovcar3 and Ovcar10) cell lines. Heat map shows z-scores of relative abundance normalized to the mean of each individual TCA cycle metabolite. Significant metabolites are labeled in green. Adjusted p-value (FDR) <0.05. (B) Differentially expressed transcripts in 5 parental fallopian tube (FT282, FT4-Tag, FT6-Tag, FT33-Tag, and PFTE4) and 4 HGSC (HeyA8, Ovcar5, Ovcar3, and Ovcar10) cell lines. Heat map shows z-scores of expression levels relative to B2M of 27 TCA cycle enzymes. Significant transcripts are labeled in green. Adjusted p-value (FDR) <0.05 (C) Differentially expressed transcripts of 27 TCA cycle enzymes. Heat map shows fold change z-scores of indicated HGSC cell lines cultured in ultra-low attachment and adherent conditions. Significant transcripts are labeled in green. Adjusted p-value (FDR) <0.001 (D) Kaplan-Meier curve for disease progression-free survival of ovarian cancer patients classified according to IDH1 expression level. Ovarian cancer patients were filtered for a serous histosubtype and TP53 mutation. Estimates of the hazard ration (HR) and logrank p-values are indicated. (E) Immunoblot analysis of IDH1 comparing normal human fallopian tube and HGSC patient tissue lysates. Vinculin was used as a loading control. (F) IDH1 protein expression normalized to vinculin and quantified using ImageJ. Data represent mean ± SD. *p<0.03.
    Figure Legend Snippet: (A) LC/MS comparison of 2 fallopian tube (FT282 and FT4-Tag) and 2 HGSC (Ovcar3 and Ovcar10) cell lines. Heat map shows z-scores of relative abundance normalized to the mean of each individual TCA cycle metabolite. Significant metabolites are labeled in green. Adjusted p-value (FDR) <0.05. (B) Differentially expressed transcripts in 5 parental fallopian tube (FT282, FT4-Tag, FT6-Tag, FT33-Tag, and PFTE4) and 4 HGSC (HeyA8, Ovcar5, Ovcar3, and Ovcar10) cell lines. Heat map shows z-scores of expression levels relative to B2M of 27 TCA cycle enzymes. Significant transcripts are labeled in green. Adjusted p-value (FDR) <0.05 (C) Differentially expressed transcripts of 27 TCA cycle enzymes. Heat map shows fold change z-scores of indicated HGSC cell lines cultured in ultra-low attachment and adherent conditions. Significant transcripts are labeled in green. Adjusted p-value (FDR) <0.001 (D) Kaplan-Meier curve for disease progression-free survival of ovarian cancer patients classified according to IDH1 expression level. Ovarian cancer patients were filtered for a serous histosubtype and TP53 mutation. Estimates of the hazard ration (HR) and logrank p-values are indicated. (E) Immunoblot analysis of IDH1 comparing normal human fallopian tube and HGSC patient tissue lysates. Vinculin was used as a loading control. (F) IDH1 protein expression normalized to vinculin and quantified using ImageJ. Data represent mean ± SD. *p<0.03.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy, Labeling, Expressing, Cell Culture, Mutagenesis, Western Blot

    (A-D) Ovcar3 cells were treated with either DMSO or 15 μM GSK864 (IDH1i) for 7 days. (A) BrdU incorporation. One of three experiments shown. (B) Quantification of (A). Data represent mean ± SD. *p<0.0001 (C) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (D) Quantification of (C). Data represent mean ± SD. *p<0.0001 (E-J) Ovcar3 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control. Cells were selected with puromycin for 7 days. (E) Immunoblot analysis of IDH1. Vinculin was used as a loading control. One of 5 experiments is shown. (F) BrdU incorporation. One of three experiments shown. (G) Quantification of (F). Data represent mean ± SD. *p<0.0001 (H) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (I) Quantification of (H). Data represent mean ± SD. *p<0.0001 (J) Immunoblot analysis of IDH1 and cyclin A. β-Actin was used as a loading control. One of three experiments shown. (K) Immunoblot of Ovcar3 cells cultured in ULA conditions and treated with 15 μM GSK864 (IDH1i) for cyclin A. β-Actin was used as a loading control. One of three experiments shown.
    Figure Legend Snippet: (A-D) Ovcar3 cells were treated with either DMSO or 15 μM GSK864 (IDH1i) for 7 days. (A) BrdU incorporation. One of three experiments shown. (B) Quantification of (A). Data represent mean ± SD. *p<0.0001 (C) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (D) Quantification of (C). Data represent mean ± SD. *p<0.0001 (E-J) Ovcar3 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control. Cells were selected with puromycin for 7 days. (E) Immunoblot analysis of IDH1. Vinculin was used as a loading control. One of 5 experiments is shown. (F) BrdU incorporation. One of three experiments shown. (G) Quantification of (F). Data represent mean ± SD. *p<0.0001 (H) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (I) Quantification of (H). Data represent mean ± SD. *p<0.0001 (J) Immunoblot analysis of IDH1 and cyclin A. β-Actin was used as a loading control. One of three experiments shown. (K) Immunoblot of Ovcar3 cells cultured in ULA conditions and treated with 15 μM GSK864 (IDH1i) for cyclin A. β-Actin was used as a loading control. One of three experiments shown.

    Techniques Used: BrdU Incorporation Assay, Staining, Infection, Expressing, Plasmid Preparation, Western Blot, Cell Culture

    (A-D) Ovcar10 cells were treated with either DMSO or 15μM GSK864 (IDH1i) for 7 days. (A) BrdU incorporation. One of three experiments shown. (B) Quantification of (A). Data represent mean ± SD. *p<0.0001 (C) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (D) Quantification of (C). Data represent mean ± SD. *p<0.0001 (E-I) Ovcar10 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control. Cells were selected with puromycin for 7 days. (E) Immunoblot analysis of IDH1. β-Actin was used as a loading control. One of 5 experiments is shown. (F) BrdU incorporation. One of three experiments shown. (G) Quantification of (G). Data represent mean ± SD. *p<0.0001 (H) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (I) Quantification of (H). Data represent mean ± SD. *p<0.0001
    Figure Legend Snippet: (A-D) Ovcar10 cells were treated with either DMSO or 15μM GSK864 (IDH1i) for 7 days. (A) BrdU incorporation. One of three experiments shown. (B) Quantification of (A). Data represent mean ± SD. *p<0.0001 (C) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (D) Quantification of (C). Data represent mean ± SD. *p<0.0001 (E-I) Ovcar10 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control. Cells were selected with puromycin for 7 days. (E) Immunoblot analysis of IDH1. β-Actin was used as a loading control. One of 5 experiments is shown. (F) BrdU incorporation. One of three experiments shown. (G) Quantification of (G). Data represent mean ± SD. *p<0.0001 (H) Colony formation. Cells were seeded at equal densities and 10 days later stained with 0.05% crystal violet. One of three experiments is shown. (I) Quantification of (H). Data represent mean ± SD. *p<0.0001

    Techniques Used: BrdU Incorporation Assay, Staining, Infection, Expressing, Plasmid Preparation, Western Blot

    (A&B) Ovcar10 cells were infected with lentivirus hairpins two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in puromycin for 7 days. (A) 7AAD flow cytometry analysis. One of three experiments is shown. *p<0.0005 (B) Brightfield image of cell morphology. (C) Ovcar10 cells were infected with lentivirus hairpins two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in puromycin for 7 days. qRT-PCR analysis of SASP gene expression (IL6, IL8, IL1α, IL1β) . Etoposide (10μM) was used as a positive control. One of three experiments is shown. Data represent mean ± SD. *p<0.0001 (D-G) Ovcar10 cells were treated with either DMSO or 15μM GSK864 (IDH1i) for 7 days. (D) SA-β-Gal activity. One of three experiments is shown. (E) Quantification of (D). Data represent mean ± SD. *p<0.002. (F) PML body foci. One of three experiments is shown. (G) Quantification of (F). Data represent mean ± SD. *p<0.0001 (H-L) Ovcar10 cells were infected with lentivirus hairpins two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in puromycin for 7 days. (H) SA-β-Gal activity. One of five experiments is shown. (I) Quantification of (H). Data represent mean ± SD. *p<0.0001 (J) Immunoblot analysis of IDH1 and lamin B1. One of three experiments is shown. (K) PML body foci. One of three experiments is shown. (IDH1i) for 4 days. Immunoblot analysis of lamin B1. One of two experiments is shown.
    Figure Legend Snippet: (A&B) Ovcar10 cells were infected with lentivirus hairpins two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in puromycin for 7 days. (A) 7AAD flow cytometry analysis. One of three experiments is shown. *p<0.0005 (B) Brightfield image of cell morphology. (C) Ovcar10 cells were infected with lentivirus hairpins two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in puromycin for 7 days. qRT-PCR analysis of SASP gene expression (IL6, IL8, IL1α, IL1β) . Etoposide (10μM) was used as a positive control. One of three experiments is shown. Data represent mean ± SD. *p<0.0001 (D-G) Ovcar10 cells were treated with either DMSO or 15μM GSK864 (IDH1i) for 7 days. (D) SA-β-Gal activity. One of three experiments is shown. (E) Quantification of (D). Data represent mean ± SD. *p<0.002. (F) PML body foci. One of three experiments is shown. (G) Quantification of (F). Data represent mean ± SD. *p<0.0001 (H-L) Ovcar10 cells were infected with lentivirus hairpins two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in puromycin for 7 days. (H) SA-β-Gal activity. One of five experiments is shown. (I) Quantification of (H). Data represent mean ± SD. *p<0.0001 (J) Immunoblot analysis of IDH1 and lamin B1. One of three experiments is shown. (K) PML body foci. One of three experiments is shown. (IDH1i) for 4 days. Immunoblot analysis of lamin B1. One of two experiments is shown.

    Techniques Used: Infection, Plasmid Preparation, Flow Cytometry, Quantitative RT-PCR, Expressing, Positive Control, Activity Assay, Western Blot

    (A-E) Ovcar3 cells were treated with either DMSO or 15μM GSK864 (IDH1i) for 7 days. (A) SA-β-Gal activity. One of three experiments is shown. (B) Quantification of (A). Data represent mean ± SD. *p<0.0001 (C) Immunoblot analysis of lamin B1. One of three experiments is shown. (D) PML body foci. One of three experiments is shown. (E) Quantification of (D). Data represent mean ± SD. *p<0.0001 (F-J) Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in 3μg/mL puromycin for 7 days. (F) SA-β-Gal activity. One of three experiments is shown. (G) Quantification of (F). Data represent mean ± SD. *p<0.0001 (H) Immunoblot analysis of IDH1 and lamin B1. One of three experiments is shown. (I) PML body foci. One of three experiments is shown. (J) Quantification of (I). Data represent mean ± SD. *p<0.0005 (K) qRT-PCR analysis of IDH1 and LMNB1 (encoding Lamin B1) in Ovcar3 shIDH1 ULA. RNA was collected four days after infection. Data represent mean ± SD. *p<0.0002 (L-O) Ovcar3 cells were infected with lentivirus expressing one shRNA targeting IDH1 (#1). Empty vector was used as a control. Cells were selected in 3μg/mL puromycin for 7 days. Exogenous αKG (1mM) was added to the indicated cells. (L) SA-β-Gal activity of shIDH1 with and without αKG. One of three experiments is shown. (M)Quantification of (L). Data represent mean ± SD. *p<0.0001 (N) Colony formation of shIDH1 with and without αKG. One of three experiments is shown. (O) Quantification of (N). Data represent mean ± SD. *p<0.002
    Figure Legend Snippet: (A-E) Ovcar3 cells were treated with either DMSO or 15μM GSK864 (IDH1i) for 7 days. (A) SA-β-Gal activity. One of three experiments is shown. (B) Quantification of (A). Data represent mean ± SD. *p<0.0001 (C) Immunoblot analysis of lamin B1. One of three experiments is shown. (D) PML body foci. One of three experiments is shown. (E) Quantification of (D). Data represent mean ± SD. *p<0.0001 (F-J) Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting IDH1. Empty vector was used as a control. Cells were selected in 3μg/mL puromycin for 7 days. (F) SA-β-Gal activity. One of three experiments is shown. (G) Quantification of (F). Data represent mean ± SD. *p<0.0001 (H) Immunoblot analysis of IDH1 and lamin B1. One of three experiments is shown. (I) PML body foci. One of three experiments is shown. (J) Quantification of (I). Data represent mean ± SD. *p<0.0005 (K) qRT-PCR analysis of IDH1 and LMNB1 (encoding Lamin B1) in Ovcar3 shIDH1 ULA. RNA was collected four days after infection. Data represent mean ± SD. *p<0.0002 (L-O) Ovcar3 cells were infected with lentivirus expressing one shRNA targeting IDH1 (#1). Empty vector was used as a control. Cells were selected in 3μg/mL puromycin for 7 days. Exogenous αKG (1mM) was added to the indicated cells. (L) SA-β-Gal activity of shIDH1 with and without αKG. One of three experiments is shown. (M)Quantification of (L). Data represent mean ± SD. *p<0.0001 (N) Colony formation of shIDH1 with and without αKG. One of three experiments is shown. (O) Quantification of (N). Data represent mean ± SD. *p<0.002

    Techniques Used: Activity Assay, Western Blot, Infection, Expressing, Plasmid Preparation, Quantitative RT-PCR, shRNA

    (A) Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Cells were selected with puromycin for 7 days. Flow cytometry analysis of ROS using DHE as a marker. Cisplatin was used as a positive control and DMF was used as a negative control. Empty vector (EV) was also used as a negative control. One of three experiments is shown. Data represent mean ± SD. *p<0.0001 (B) Ovcar10 cells were infected with two independent hairpins targeting IDH1. Cells were selected with puromycin for 7 days. 500μM of n-acetyl cysteine (NAC) or vehicle control (water) was added to cells. Cells were seeded in 6-well plates and stained with crystal violet. Quantification O.D. 590 was determined. One of two experiments is shown. Data represent mean ± SD. *p<0.005 (C) Ovcar3 cells were infected with two independent hairpins targeting IDH1. Cells were selected with puromycin for 7 days. Immunofluorescence quantification of 53BP1 and γH2AX. Cisplatin (1μM) was used as a positive control. One of three experiments is shown. Data represent mean ± SD. *p<0.02.
    Figure Legend Snippet: (A) Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Cells were selected with puromycin for 7 days. Flow cytometry analysis of ROS using DHE as a marker. Cisplatin was used as a positive control and DMF was used as a negative control. Empty vector (EV) was also used as a negative control. One of three experiments is shown. Data represent mean ± SD. *p<0.0001 (B) Ovcar10 cells were infected with two independent hairpins targeting IDH1. Cells were selected with puromycin for 7 days. 500μM of n-acetyl cysteine (NAC) or vehicle control (water) was added to cells. Cells were seeded in 6-well plates and stained with crystal violet. Quantification O.D. 590 was determined. One of two experiments is shown. Data represent mean ± SD. *p<0.005 (C) Ovcar3 cells were infected with two independent hairpins targeting IDH1. Cells were selected with puromycin for 7 days. Immunofluorescence quantification of 53BP1 and γH2AX. Cisplatin (1μM) was used as a positive control. One of three experiments is shown. Data represent mean ± SD. *p<0.02.

    Techniques Used: Infection, Flow Cytometry, Marker, Positive Control, Negative Control, Plasmid Preparation, Staining, Immunofluorescence

    (A&B) Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Empty vector was used as a control. Cells were selected with puromycin for 7 days. (A) Chromatin immunoprecipitation qPCR analysis of H3K9me2 at the loci of PCNA and MCM3 . Data is normalized to a gene desert region. One of two experiments is shown. Data represent mean ± SD. *p<0.02 (B) RT-qPCR analysis PCNA and MCM3 expression. B2M was used as a reference gene. One of three experiments is shown. Data represent mean ± SD. *p<0.008
    Figure Legend Snippet: (A&B) Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Empty vector was used as a control. Cells were selected with puromycin for 7 days. (A) Chromatin immunoprecipitation qPCR analysis of H3K9me2 at the loci of PCNA and MCM3 . Data is normalized to a gene desert region. One of two experiments is shown. Data represent mean ± SD. *p<0.02 (B) RT-qPCR analysis PCNA and MCM3 expression. B2M was used as a reference gene. One of three experiments is shown. Data represent mean ± SD. *p<0.008

    Techniques Used: Infection, Plasmid Preparation, Chromatin Immunoprecipitation, Quantitative RT-PCR, Expressing

    idh1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc idh1
    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , <t>IDH1</t> and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.
    Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/idh1/product/Cell Signaling Technology Inc
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    1) Product Images from "An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis"

    Article Title: An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.101326

    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , IDH1 and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.
    Figure Legend Snippet: ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , IDH1 and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.

    Techniques Used: Expressing, Immunofluorescence, Immunohistochemistry, Labeling

    ( A ) The expression levels of AVEN, IDH1 and LOX proteins were evaluated by western blotting in cultured keratinocytes transduced by the miR-30a lentivirus construction after doxycycline treatment. ( B ) Immuno-fluorescent staining of AVEN and IDH1 and immunohistochemical staining of LOX in reconstructed epidermis overexpressing or not miR-30a after doxycycline treatment. Counterstaining was performed with DAPI and polycarbonate membrane position is indicated by a dotted line. Representative photographs of 3 independent replicates were shown. Scale bar = 25 μm. Right panels: quantification of the IDH1, LOX or AVEN labeled area in the REs Results are mean +/− SD from three independent samples. ****P<0,0001, ***P<0,001 ( C ) The alignment of miR-30a putative binding sites in human AVEN , IDH1 or LOX 3′-UTR region have been schematized to show complementary pairing of miR-30a with AVEN , IDH1 or LOX wild-type (WT) and mutant (Mut) 3′-UTR constructs. Transduced keratinocytes were transfected with WT or Mut reporter constructs. Luciferase intensities were normalized to β-galactosidase level. Results are mean +/− SD from three independent samples. *P<0,05, **P<0,01, ***P<0,001, ****P<0,0001.
    Figure Legend Snippet: ( A ) The expression levels of AVEN, IDH1 and LOX proteins were evaluated by western blotting in cultured keratinocytes transduced by the miR-30a lentivirus construction after doxycycline treatment. ( B ) Immuno-fluorescent staining of AVEN and IDH1 and immunohistochemical staining of LOX in reconstructed epidermis overexpressing or not miR-30a after doxycycline treatment. Counterstaining was performed with DAPI and polycarbonate membrane position is indicated by a dotted line. Representative photographs of 3 independent replicates were shown. Scale bar = 25 μm. Right panels: quantification of the IDH1, LOX or AVEN labeled area in the REs Results are mean +/− SD from three independent samples. ****P<0,0001, ***P<0,001 ( C ) The alignment of miR-30a putative binding sites in human AVEN , IDH1 or LOX 3′-UTR region have been schematized to show complementary pairing of miR-30a with AVEN , IDH1 or LOX wild-type (WT) and mutant (Mut) 3′-UTR constructs. Transduced keratinocytes were transfected with WT or Mut reporter constructs. Luciferase intensities were normalized to β-galactosidase level. Results are mean +/− SD from three independent samples. *P<0,05, **P<0,01, ***P<0,001, ****P<0,0001.

    Techniques Used: Expressing, Western Blot, Cell Culture, Staining, Immunohistochemical staining, Labeling, Binding Assay, Mutagenesis, Construct, Transfection, Luciferase

    idh1  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc idh1
    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , <t>IDH1</t> and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.
    Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/idh1/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    idh1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis"

    Article Title: An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.101326

    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , IDH1 and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.
    Figure Legend Snippet: ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , IDH1 and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.

    Techniques Used: Expressing, Immunofluorescence, Immunohistochemistry, Labeling

    ( A ) The expression levels of AVEN, IDH1 and LOX proteins were evaluated by western blotting in cultured keratinocytes transduced by the miR-30a lentivirus construction after doxycycline treatment. ( B ) Immuno-fluorescent staining of AVEN and IDH1 and immunohistochemical staining of LOX in reconstructed epidermis overexpressing or not miR-30a after doxycycline treatment. Counterstaining was performed with DAPI and polycarbonate membrane position is indicated by a dotted line. Representative photographs of 3 independent replicates were shown. Scale bar = 25 μm. Right panels: quantification of the IDH1, LOX or AVEN labeled area in the REs Results are mean +/− SD from three independent samples. ****P<0,0001, ***P<0,001 ( C ) The alignment of miR-30a putative binding sites in human AVEN , IDH1 or LOX 3′-UTR region have been schematized to show complementary pairing of miR-30a with AVEN , IDH1 or LOX wild-type (WT) and mutant (Mut) 3′-UTR constructs. Transduced keratinocytes were transfected with WT or Mut reporter constructs. Luciferase intensities were normalized to β-galactosidase level. Results are mean +/− SD from three independent samples. *P<0,05, **P<0,01, ***P<0,001, ****P<0,0001.
    Figure Legend Snippet: ( A ) The expression levels of AVEN, IDH1 and LOX proteins were evaluated by western blotting in cultured keratinocytes transduced by the miR-30a lentivirus construction after doxycycline treatment. ( B ) Immuno-fluorescent staining of AVEN and IDH1 and immunohistochemical staining of LOX in reconstructed epidermis overexpressing or not miR-30a after doxycycline treatment. Counterstaining was performed with DAPI and polycarbonate membrane position is indicated by a dotted line. Representative photographs of 3 independent replicates were shown. Scale bar = 25 μm. Right panels: quantification of the IDH1, LOX or AVEN labeled area in the REs Results are mean +/− SD from three independent samples. ****P<0,0001, ***P<0,001 ( C ) The alignment of miR-30a putative binding sites in human AVEN , IDH1 or LOX 3′-UTR region have been schematized to show complementary pairing of miR-30a with AVEN , IDH1 or LOX wild-type (WT) and mutant (Mut) 3′-UTR constructs. Transduced keratinocytes were transfected with WT or Mut reporter constructs. Luciferase intensities were normalized to β-galactosidase level. Results are mean +/− SD from three independent samples. *P<0,05, **P<0,01, ***P<0,001, ****P<0,0001.

    Techniques Used: Expressing, Western Blot, Cell Culture, Staining, Immunohistochemical staining, Labeling, Binding Assay, Mutagenesis, Construct, Transfection, Luciferase

    anti idh1 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti idh1 antibody
    Anti Idh1 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti idh1 antibody/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
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    Cell Signaling Technology Inc rabbit igg anti idh1
    <t>IDH1</t> catalyzes the conversion of isocitrate to αKG.
    Rabbit Igg Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phycoerythrin pe conjugated anti idh1
    Proteome-transcriptome correlation in human hematopoietic stem and progenitor cell subpopulations. ( A ) Nonsupervised hierarchical clustering (Euclidean distance) heatmap of the intensity for the transcripts identified in HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs (shades of red) isolated from five different HSPC donors (shades of blue). The transcript intensities are centered and scaled and depicted in color shades from red to blue. The transcripts with missing transcript intensity values in all samples were removed because they could not be handled by the clustering algorithm. Remaining missing transcript intensity values are shown in white. Clustering was observed mostly according to cell type, not according to donor. ( B ) GO enrichment analysis showed good alignment of protein and mRNA data. GSEA was performed for ranked mRNA and protein lists using GO processes from the Gene Ontology Consortium database as gene sets. Shown are normalized enrichment scores for the individual gene sets. Significantly up-regulated gene sets are marked in blue color; significantly downregulated gene sets are marked in red color. Significance was defined as FDR < 0.25, specific cell subpopulations were compared with the average of the remaining three cell types, and log2(fold change) was used as ranking criterion. Empty fields mean that no enrichment could be calculated. Abbreviations: MEGA, megakaryocyte; MAPK, mitogen-activated protein kinase; PI3K, phosphoinositide-3-kinase; PLC, phospholipase C. ( C ) Correlation between proteomics and transcriptomics data for HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs. Dots are depicted in red when the FDR was below 0.01 both for protein and RNA data, orange when FDR < 0.01 for protein data, purple when FDR < 0.01 for RNA data, and gray when FDR ≥ 0.01 for both protein and RNA data. ( D ) Network analysis of significantly up-regulated proteins with concomitant significantly downregulated mRNA in HSCs/MPPs. Two clusters were especially prominent, including the snoRNPs and telomerase complex proteins GAR1, DKC1, NOP10, NHP2, and the quiescence-inducing NAD(P)H-producing proteins <t>IDH1,</t> IDH3A, and IDH3B. HSCs/MPPs were compared with the average of the other three subpopulations; cutoffs were set at FDR < 0.01 for protein and RNA data. Colors depict GO terms found enriched in B .
    Phycoerythrin Pe Conjugated Anti Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc idh1
    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , <t>IDH1</t> and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.
    Idh1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/idh1/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc anti idh1 antibody
    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , <t>IDH1</t> and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.
    Anti Idh1 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti idh1 antibody/product/Cell Signaling Technology Inc
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    Image Search Results


    IDH1 catalyzes the conversion of isocitrate to αKG.

    Journal: The Analyst

    Article Title: A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1

    doi: 10.1039/d0an00174k

    Figure Lengend Snippet: IDH1 catalyzes the conversion of isocitrate to αKG.

    Article Snippet: The membrane was then blocked using Odyssey® TBS blocking buffer (LI-COR, Lincoln, NE) at room temperature for 1 hour and incubated overnight with rabbit IgG anti-IDH1 (Cell Signaling Technology, Danvers, MA) at a 1 : 1000 dilution and mouse IgG1 anti-HSP70 (BD Biosciences, Franklin Lanes, NJ) at a 1 : 2000 dilution at 4 °C.

    Techniques:

    (a) Scheme for the preparation and analysis of recombinant IDH1 samples. (b) αKG concentrations measured in the 384-sample biochemical screen (7 samples did not produce either 12C or 13C αKG capture).

    Journal: The Analyst

    Article Title: A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1

    doi: 10.1039/d0an00174k

    Figure Lengend Snippet: (a) Scheme for the preparation and analysis of recombinant IDH1 samples. (b) αKG concentrations measured in the 384-sample biochemical screen (7 samples did not produce either 12C or 13C αKG capture).

    Article Snippet: The membrane was then blocked using Odyssey® TBS blocking buffer (LI-COR, Lincoln, NE) at room temperature for 1 hour and incubated overnight with rabbit IgG anti-IDH1 (Cell Signaling Technology, Danvers, MA) at a 1 : 1000 dilution and mouse IgG1 anti-HSP70 (BD Biosciences, Franklin Lanes, NJ) at a 1 : 2000 dilution at 4 °C.

    Techniques: Recombinant

    IDH1 expression level as determined with western blot and αKG concentration as measured with SAMDI-MS for untreated cells and cells treated with transfection agent, control sequence siRNA and IDH1-targeting siRNA. All data are expressed as mean ± standard deviation with n = 2 experimental replicates.

    Journal: The Analyst

    Article Title: A high-throughput SAMDI-mass spectrometry assay for isocitrate dehydrogenase 1

    doi: 10.1039/d0an00174k

    Figure Lengend Snippet: IDH1 expression level as determined with western blot and αKG concentration as measured with SAMDI-MS for untreated cells and cells treated with transfection agent, control sequence siRNA and IDH1-targeting siRNA. All data are expressed as mean ± standard deviation with n = 2 experimental replicates.

    Article Snippet: The membrane was then blocked using Odyssey® TBS blocking buffer (LI-COR, Lincoln, NE) at room temperature for 1 hour and incubated overnight with rabbit IgG anti-IDH1 (Cell Signaling Technology, Danvers, MA) at a 1 : 1000 dilution and mouse IgG1 anti-HSP70 (BD Biosciences, Franklin Lanes, NJ) at a 1 : 2000 dilution at 4 °C.

    Techniques: Expressing, Western Blot, Concentration Assay, Transfection, Sequencing, Standard Deviation

    Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting IDH1 or treated with 15μM GSK864 (IDH1i). Empty vector or DMSO was used as a control, respectively.

    Journal: Molecular cancer research : MCR

    Article Title: Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer

    doi: 10.1158/1541-7786.MCR-18-1233

    Figure Lengend Snippet: Ovcar3 cells were infected with lentivirus expressing two independent shRNAs targeting IDH1 or treated with 15μM GSK864 (IDH1i). Empty vector or DMSO was used as a control, respectively.

    Article Snippet: The following antibodies were obtained from the indicated suppliers: rabbit anti-IDH1 (Cell Signaling), rabbit anti-Lamin B1 (Abcam), rabbit anti-Cyclin A (Abcam), mouse anti-PCNA (Cell Signaling), mouse anti-MCM3 (Santa Cruz Biotechnologies), mouse anti-Vinculin (Sigma-Aldrich), mouse anti-Beta Actin (Sigma-Aldrich), rat anti-BrdU (Abcam), mouse anti-PML (Santa Cruz Biotechnologies), mouse anti-γH2AX (EMD Millipore), rabbit anti-53BP1 (Bethyl), Fluorescein donkey anti-rat IgG (Jackson ImmunoResearch), Cy™3 donkey anti-mouse (Jackson ImmunoResearch).

    Techniques: Infection, Expressing, Plasmid Preparation

    (A-F) Ovcar3 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control.

    Journal: Molecular cancer research : MCR

    Article Title: Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer

    doi: 10.1158/1541-7786.MCR-18-1233

    Figure Lengend Snippet: (A-F) Ovcar3 cells were infected with lentivirus expressing two independent short hairpin RNAs (shRNAs) targeting IDH1 (shIDH1). Empty vector was used as a control.

    Article Snippet: The following antibodies were obtained from the indicated suppliers: rabbit anti-IDH1 (Cell Signaling), rabbit anti-Lamin B1 (Abcam), rabbit anti-Cyclin A (Abcam), mouse anti-PCNA (Cell Signaling), mouse anti-MCM3 (Santa Cruz Biotechnologies), mouse anti-Vinculin (Sigma-Aldrich), mouse anti-Beta Actin (Sigma-Aldrich), rat anti-BrdU (Abcam), mouse anti-PML (Santa Cruz Biotechnologies), mouse anti-γH2AX (EMD Millipore), rabbit anti-53BP1 (Bethyl), Fluorescein donkey anti-rat IgG (Jackson ImmunoResearch), Cy™3 donkey anti-mouse (Jackson ImmunoResearch).

    Techniques: Infection, Expressing, Plasmid Preparation

    Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Empty vector was used as a control.

    Journal: Molecular cancer research : MCR

    Article Title: Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer

    doi: 10.1158/1541-7786.MCR-18-1233

    Figure Lengend Snippet: Ovcar3 and Ovcar10 cells were infected with two independent hairpins targeting IDH1. Empty vector was used as a control.

    Article Snippet: The following antibodies were obtained from the indicated suppliers: rabbit anti-IDH1 (Cell Signaling), rabbit anti-Lamin B1 (Abcam), rabbit anti-Cyclin A (Abcam), mouse anti-PCNA (Cell Signaling), mouse anti-MCM3 (Santa Cruz Biotechnologies), mouse anti-Vinculin (Sigma-Aldrich), mouse anti-Beta Actin (Sigma-Aldrich), rat anti-BrdU (Abcam), mouse anti-PML (Santa Cruz Biotechnologies), mouse anti-γH2AX (EMD Millipore), rabbit anti-53BP1 (Bethyl), Fluorescein donkey anti-rat IgG (Jackson ImmunoResearch), Cy™3 donkey anti-mouse (Jackson ImmunoResearch).

    Techniques: Infection, Plasmid Preparation

    Genetic mutations in  isocitrate dehydrogenase  (IDH)1/2

    Journal: Cancer Science

    Article Title: Pharmacological characterization of TQ 05310, a potent inhibitor of isocitrate dehydrogenase 2 R140Q and R172K mutants

    doi: 10.1111/cas.14152

    Figure Lengend Snippet: Genetic mutations in isocitrate dehydrogenase (IDH)1/2

    Article Snippet: Monoclonal antibodies against IDH1 and Myc‐tag were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Mutagenesis

    Journal: Cell Metabolism

    Article Title: Polyamines and eIF5A Hypusination Modulate Mitochondrial Respiration and Macrophage Activation

    doi: 10.1016/j.cmet.2019.05.003

    Figure Lengend Snippet:

    Article Snippet: Anti-IDH1 , Cell signalling , D2H1.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Construct, Plasmid Preparation, Sequencing, Clone Assay, Software

    Proteome-transcriptome correlation in human hematopoietic stem and progenitor cell subpopulations. ( A ) Nonsupervised hierarchical clustering (Euclidean distance) heatmap of the intensity for the transcripts identified in HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs (shades of red) isolated from five different HSPC donors (shades of blue). The transcript intensities are centered and scaled and depicted in color shades from red to blue. The transcripts with missing transcript intensity values in all samples were removed because they could not be handled by the clustering algorithm. Remaining missing transcript intensity values are shown in white. Clustering was observed mostly according to cell type, not according to donor. ( B ) GO enrichment analysis showed good alignment of protein and mRNA data. GSEA was performed for ranked mRNA and protein lists using GO processes from the Gene Ontology Consortium database as gene sets. Shown are normalized enrichment scores for the individual gene sets. Significantly up-regulated gene sets are marked in blue color; significantly downregulated gene sets are marked in red color. Significance was defined as FDR < 0.25, specific cell subpopulations were compared with the average of the remaining three cell types, and log2(fold change) was used as ranking criterion. Empty fields mean that no enrichment could be calculated. Abbreviations: MEGA, megakaryocyte; MAPK, mitogen-activated protein kinase; PI3K, phosphoinositide-3-kinase; PLC, phospholipase C. ( C ) Correlation between proteomics and transcriptomics data for HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs. Dots are depicted in red when the FDR was below 0.01 both for protein and RNA data, orange when FDR < 0.01 for protein data, purple when FDR < 0.01 for RNA data, and gray when FDR ≥ 0.01 for both protein and RNA data. ( D ) Network analysis of significantly up-regulated proteins with concomitant significantly downregulated mRNA in HSCs/MPPs. Two clusters were especially prominent, including the snoRNPs and telomerase complex proteins GAR1, DKC1, NOP10, NHP2, and the quiescence-inducing NAD(P)H-producing proteins IDH1, IDH3A, and IDH3B. HSCs/MPPs were compared with the average of the other three subpopulations; cutoffs were set at FDR < 0.01 for protein and RNA data. Colors depict GO terms found enriched in B .

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Sensitive Quantitative Proteomics of Human Hematopoietic Stem and Progenitor Cells by Data-independent Acquisition Mass Spectrometry

    doi: 10.1074/mcp.TIR119.001431

    Figure Lengend Snippet: Proteome-transcriptome correlation in human hematopoietic stem and progenitor cell subpopulations. ( A ) Nonsupervised hierarchical clustering (Euclidean distance) heatmap of the intensity for the transcripts identified in HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs (shades of red) isolated from five different HSPC donors (shades of blue). The transcript intensities are centered and scaled and depicted in color shades from red to blue. The transcripts with missing transcript intensity values in all samples were removed because they could not be handled by the clustering algorithm. Remaining missing transcript intensity values are shown in white. Clustering was observed mostly according to cell type, not according to donor. ( B ) GO enrichment analysis showed good alignment of protein and mRNA data. GSEA was performed for ranked mRNA and protein lists using GO processes from the Gene Ontology Consortium database as gene sets. Shown are normalized enrichment scores for the individual gene sets. Significantly up-regulated gene sets are marked in blue color; significantly downregulated gene sets are marked in red color. Significance was defined as FDR < 0.25, specific cell subpopulations were compared with the average of the remaining three cell types, and log2(fold change) was used as ranking criterion. Empty fields mean that no enrichment could be calculated. Abbreviations: MEGA, megakaryocyte; MAPK, mitogen-activated protein kinase; PI3K, phosphoinositide-3-kinase; PLC, phospholipase C. ( C ) Correlation between proteomics and transcriptomics data for HSCs/MPPs (referred to as HSCs), CMPs, GMPs, and MEPs. Dots are depicted in red when the FDR was below 0.01 both for protein and RNA data, orange when FDR < 0.01 for protein data, purple when FDR < 0.01 for RNA data, and gray when FDR ≥ 0.01 for both protein and RNA data. ( D ) Network analysis of significantly up-regulated proteins with concomitant significantly downregulated mRNA in HSCs/MPPs. Two clusters were especially prominent, including the snoRNPs and telomerase complex proteins GAR1, DKC1, NOP10, NHP2, and the quiescence-inducing NAD(P)H-producing proteins IDH1, IDH3A, and IDH3B. HSCs/MPPs were compared with the average of the other three subpopulations; cutoffs were set at FDR < 0.01 for protein and RNA data. Colors depict GO terms found enriched in B .

    Article Snippet: For FACS analysis of intracellular isocitrate dehydrogenase 1 (IDH1) expression, the IntraPrep Leukocytic Permeabilization Reagent Kit (Beckman Coulter, Brea, CA) was used together with phycoerythrin (PE)-conjugated anti-IDH1, D2H1 (Cell Signaling Technology, Danvers, MA) and PE-conjugated isotype control, DA1E (Cell Signaling Technology).

    Techniques: Isolation

    ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , IDH1 and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.

    Journal: Aging (Albany NY)

    Article Title: An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis

    doi: 10.18632/aging.101326

    Figure Lengend Snippet: ( A ) A list of genes potentially targeted by miR-30a according to transcriptome data available in the GEO database. The number of conserved miR-30a MRE is indicated for each gene. The potential expression of these genes in human epidermis (according to the Human Protein Atlas database) is indicated. Five genes exhibiting a least one miR-30a conserved MRE and expressed in human epidermis, were analyzed by QPCR in RNA samples from skin equivalent (SE) model mimicking aging. The expression profile is indicated for these 5 genes. ( B ) The expression levels of AVEN , IDH1 and LOX were evaluated by QPCR in RNA samples from SEs mimicking aging with a long time of culture from day 35 (D35) to day 100 (D100). Results are mean +/− SD from three independent samples. *P< 0,05, ****P<0,0001. ( C ) Expression of AVEN, IDH1 or LOX in young or old human skin sections assessed by immunofluorescence for AVEN and IDH1 and by immunohistochemistry for LOX. (D) Quantification of the AVEN, IDH1 and LOX labeled area in the epidermis from young or old human skin sections. Results are mean +/− SD from three independent samples. ***P<0,001, **P<0,01.

    Article Snippet: The membrane was incubated overnight at 4°C in TBS-T with primary antibody specific to IDH1 (Cell Signaling Technology, Danvers, USA #D2H1, 1/1000), LOX (home-made antibody, 1/250), AVEN (Sigma, St-Louis, USA #HPA020863, 1/100) or ACTIN (Millipore, Billerica, USA #MAB 1501 Clone C4, 1/5000).

    Techniques: Expressing, Immunofluorescence, Immunohistochemistry, Labeling

    ( A ) The expression levels of AVEN, IDH1 and LOX proteins were evaluated by western blotting in cultured keratinocytes transduced by the miR-30a lentivirus construction after doxycycline treatment. ( B ) Immuno-fluorescent staining of AVEN and IDH1 and immunohistochemical staining of LOX in reconstructed epidermis overexpressing or not miR-30a after doxycycline treatment. Counterstaining was performed with DAPI and polycarbonate membrane position is indicated by a dotted line. Representative photographs of 3 independent replicates were shown. Scale bar = 25 μm. Right panels: quantification of the IDH1, LOX or AVEN labeled area in the REs Results are mean +/− SD from three independent samples. ****P<0,0001, ***P<0,001 ( C ) The alignment of miR-30a putative binding sites in human AVEN , IDH1 or LOX 3′-UTR region have been schematized to show complementary pairing of miR-30a with AVEN , IDH1 or LOX wild-type (WT) and mutant (Mut) 3′-UTR constructs. Transduced keratinocytes were transfected with WT or Mut reporter constructs. Luciferase intensities were normalized to β-galactosidase level. Results are mean +/− SD from three independent samples. *P<0,05, **P<0,01, ***P<0,001, ****P<0,0001.

    Journal: Aging (Albany NY)

    Article Title: An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis

    doi: 10.18632/aging.101326

    Figure Lengend Snippet: ( A ) The expression levels of AVEN, IDH1 and LOX proteins were evaluated by western blotting in cultured keratinocytes transduced by the miR-30a lentivirus construction after doxycycline treatment. ( B ) Immuno-fluorescent staining of AVEN and IDH1 and immunohistochemical staining of LOX in reconstructed epidermis overexpressing or not miR-30a after doxycycline treatment. Counterstaining was performed with DAPI and polycarbonate membrane position is indicated by a dotted line. Representative photographs of 3 independent replicates were shown. Scale bar = 25 μm. Right panels: quantification of the IDH1, LOX or AVEN labeled area in the REs Results are mean +/− SD from three independent samples. ****P<0,0001, ***P<0,001 ( C ) The alignment of miR-30a putative binding sites in human AVEN , IDH1 or LOX 3′-UTR region have been schematized to show complementary pairing of miR-30a with AVEN , IDH1 or LOX wild-type (WT) and mutant (Mut) 3′-UTR constructs. Transduced keratinocytes were transfected with WT or Mut reporter constructs. Luciferase intensities were normalized to β-galactosidase level. Results are mean +/− SD from three independent samples. *P<0,05, **P<0,01, ***P<0,001, ****P<0,0001.

    Article Snippet: The membrane was incubated overnight at 4°C in TBS-T with primary antibody specific to IDH1 (Cell Signaling Technology, Danvers, USA #D2H1, 1/1000), LOX (home-made antibody, 1/250), AVEN (Sigma, St-Louis, USA #HPA020863, 1/100) or ACTIN (Millipore, Billerica, USA #MAB 1501 Clone C4, 1/5000).

    Techniques: Expressing, Western Blot, Cell Culture, Staining, Immunohistochemical staining, Labeling, Binding Assay, Mutagenesis, Construct, Transfection, Luciferase