length cdna Search Results


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  • 85
    Addgene inc full length myap complementary dna cdna
    Full Length Myap Complementary Dna Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene length complementary dna cdna
    Interdependent membrane expression of cubilin and <t>amnionless.</t> ( a ) Rat cubilin (full-length and mini cubilin; 1–930) and human amnionless (full length) constructs encoded by plasmid <t>cDNA</t> used for transient transfection of cultured cells. ( b , d , f ) Non-permeabilised HEK293T, MDCK cells and RPTECs transfected with the indicated vectors were fixed and stained for membrane-targeted cubilin (red). GFP-tagged amnionless is shown in green and DAPI nuclear staining in blue. (Scale bar: 10 µm). ( c , e , g ) Plasma membrane expressions of cubilin were obtained in permeabilised HEK293T, MDCK cells and RPTECs cotransfected with amnionless. Pictures are confocal sections taken from the middle height of cells ( top ) and X-Z vertical sections ( bottom ) (Scale bar: 10 µm).
    Length Complementary Dna Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene full length human rnaseh1 complementary dna cdna
    <t>RNaseH1</t> functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. <t>DNA</t> was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P
    Full Length Human Rnaseh1 Complementary Dna Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene full length wt mps1 complementary dna
    <t>RNaseH1</t> functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. <t>DNA</t> was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P
    Full Length Wt Mps1 Complementary Dna, supplied by OriGene, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GenScript full length complementary dna cdna
    <t>RNaseH1</t> functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. <t>DNA</t> was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P
    Full Length Complementary Dna Cdna, supplied by GenScript, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Oxford Nanopore full length complementary dna cdna
    <t>RNaseH1</t> functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. <t>DNA</t> was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P
    Full Length Complementary Dna Cdna, supplied by Oxford Nanopore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher full length complementary dna cdna
    <t>RNaseH1</t> functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. <t>DNA</t> was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P
    Full Length Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa full length complementary dna cdna
    In vivo association and domain mapping of p53 and <t>EAF2</t> protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of <t>DNA</t> damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.
    Full Length Complementary Dna Cdna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery full length complementary dna cdna
    In vivo association and domain mapping of p53 and <t>EAF2</t> protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of <t>DNA</t> damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.
    Full Length Complementary Dna Cdna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    St. Jude Children's Research Hospital full length complementary dna
    In vivo association and domain mapping of p53 and <t>EAF2</t> protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of <t>DNA</t> damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.
    Full Length Complementary Dna, supplied by St. Jude Children's Research Hospital, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genewiz full length complementary dna
    In vivo association and domain mapping of p53 and <t>EAF2</t> protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of <t>DNA</t> damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.
    Full Length Complementary Dna, supplied by Genewiz, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene human full length gelsolin complementary dna cdna
    In vivo association and domain mapping of p53 and <t>EAF2</t> protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of <t>DNA</t> damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.
    Human Full Length Gelsolin Complementary Dna Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    OriGene mice full length mouse rgs14 complementary dna cdna
    Schematic diagram of the construction of <t>RGS14</t> -KO mice using the CRISPR-Cas9 method and identification of RGS14 expression. a One sgRNA targeting a region downstream of the 3′ end of exon 3 in the RGS14 mouse gene was designed and constructed. b After microinjection, a T7E1 assay indicated that four out of six pups contained cleavage products, suggesting a mixture of mutant and wild-type <t>DNA</t> templates in these mice. c Following subcloning of the PCR products, eight subclones of each mouse were sequenced. All of the subclones carried a single mutant allele, whereas two indels (#5–5, #5–6) produced frameshift mutations. Founder #5–5 was mated to a C57BL/6J mouse to obtain the F1 generation. d Gene sequencing for RGS14 expression levels in hearts from wild-type and RGS14 knockout groups. e Representative western blots for RGS14 expression levels in hearts from the RGS14 +/+ and RGS14 −/− groups
    Mice Full Length Mouse Rgs14 Complementary Dna Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vertex Pharmaceuticals full length chimeric hcv complementary dna cdna
    Schematic diagram of the construction of <t>RGS14</t> -KO mice using the CRISPR-Cas9 method and identification of RGS14 expression. a One sgRNA targeting a region downstream of the 3′ end of exon 3 in the RGS14 mouse gene was designed and constructed. b After microinjection, a T7E1 assay indicated that four out of six pups contained cleavage products, suggesting a mixture of mutant and wild-type <t>DNA</t> templates in these mice. c Following subcloning of the PCR products, eight subclones of each mouse were sequenced. All of the subclones carried a single mutant allele, whereas two indels (#5–5, #5–6) produced frameshift mutations. Founder #5–5 was mated to a C57BL/6J mouse to obtain the F1 generation. d Gene sequencing for RGS14 expression levels in hearts from wild-type and RGS14 knockout groups. e Representative western blots for RGS14 expression levels in hearts from the RGS14 +/+ and RGS14 −/− groups
    Full Length Chimeric Hcv Complementary Dna Cdna, supplied by Vertex Pharmaceuticals, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher length human anxa1 complementary dna cdna
    Schematic diagram of the construction of <t>RGS14</t> -KO mice using the CRISPR-Cas9 method and identification of RGS14 expression. a One sgRNA targeting a region downstream of the 3′ end of exon 3 in the RGS14 mouse gene was designed and constructed. b After microinjection, a T7E1 assay indicated that four out of six pups contained cleavage products, suggesting a mixture of mutant and wild-type <t>DNA</t> templates in these mice. c Following subcloning of the PCR products, eight subclones of each mouse were sequenced. All of the subclones carried a single mutant allele, whereas two indels (#5–5, #5–6) produced frameshift mutations. Founder #5–5 was mated to a C57BL/6J mouse to obtain the F1 generation. d Gene sequencing for RGS14 expression levels in hearts from wild-type and RGS14 knockout groups. e Representative western blots for RGS14 expression levels in hearts from the RGS14 +/+ and RGS14 −/− groups
    Length Human Anxa1 Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Seegene full length complementary dna
    Dual targeting of <t>DNA</t> topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. <t>glauca</t> (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA
    Full Length Complementary Dna, supplied by Seegene, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher transduction full length human nfib complementary dna cdna
    Dual targeting of <t>DNA</t> topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. <t>glauca</t> (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA
    Transduction Full Length Human Nfib Complementary Dna Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher full length tagln complementary dna
    Dual targeting of <t>DNA</t> topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. <t>glauca</t> (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA
    Full Length Tagln Complementary Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genecopoeia full length hmga2 complementary dna cdna
    Dual targeting of <t>DNA</t> topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. <t>glauca</t> (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA
    Full Length Hmga2 Complementary Dna Cdna, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    EUROIMMUN complementary dna encoding full length pla2 r isoform 1
    Dual targeting of <t>DNA</t> topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. <t>glauca</t> (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA
    Complementary Dna Encoding Full Length Pla2 R Isoform 1, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Interdependent membrane expression of cubilin and amnionless. ( a ) Rat cubilin (full-length and mini cubilin; 1–930) and human amnionless (full length) constructs encoded by plasmid cDNA used for transient transfection of cultured cells. ( b , d , f ) Non-permeabilised HEK293T, MDCK cells and RPTECs transfected with the indicated vectors were fixed and stained for membrane-targeted cubilin (red). GFP-tagged amnionless is shown in green and DAPI nuclear staining in blue. (Scale bar: 10 µm). ( c , e , g ) Plasma membrane expressions of cubilin were obtained in permeabilised HEK293T, MDCK cells and RPTECs cotransfected with amnionless. Pictures are confocal sections taken from the middle height of cells ( top ) and X-Z vertical sections ( bottom ) (Scale bar: 10 µm).

    Journal: Scientific Reports

    Article Title: Amnionless-mediated glycosylation is crucial for cell surface targeting of cubilin in renal and intestinal cells

    doi: 10.1038/s41598-018-20731-4

    Figure Lengend Snippet: Interdependent membrane expression of cubilin and amnionless. ( a ) Rat cubilin (full-length and mini cubilin; 1–930) and human amnionless (full length) constructs encoded by plasmid cDNA used for transient transfection of cultured cells. ( b , d , f ) Non-permeabilised HEK293T, MDCK cells and RPTECs transfected with the indicated vectors were fixed and stained for membrane-targeted cubilin (red). GFP-tagged amnionless is shown in green and DAPI nuclear staining in blue. (Scale bar: 10 µm). ( c , e , g ) Plasma membrane expressions of cubilin were obtained in permeabilised HEK293T, MDCK cells and RPTECs cotransfected with amnionless. Pictures are confocal sections taken from the middle height of cells ( top ) and X-Z vertical sections ( bottom ) (Scale bar: 10 µm).

    Article Snippet: Expression vectors Plasmids containing full length cDNA for wild-type human amnionless with the C-terminal mycDDK tag were purchased from OriGene Technologies (Rockville, MD).

    Techniques: Expressing, Construct, Plasmid Preparation, Transfection, Cell Culture, Staining

    RNaseH1 functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. DNA was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P

    Journal: Nature Communications

    Article Title: RNaseH1 regulates TERRA-telomeric DNA hybrids and telomere maintenance in ALT tumour cells

    doi: 10.1038/ncomms6220

    Figure Lengend Snippet: RNaseH1 functions at ALT telomeres. ( a ) Quantification of ChIP dot blot experiments performed in the indicated cell lines using antibodies against endogenous RNaseH1. DNA was first hybridized with telomeric probes and then with Alu repeat probes for specificity. Immunoprecipitated DNA is expressed as fraction of input DNA after subtraction of the background signal associated to control ChIPs using normal IgGs. Bars and error bars are averages and s.d. from three to five experiments. ( b ) Quantification of telomeric hybrids from the indicated chromosome ends and actin locus measured in S9.6 DIP experiments using cells transfected for 72 h with control siRNAs (siCtrl, set to 1) or siRNAs against RNaseH1 (siRH1a and c). Hybrids are expressed as fractions of the input material after subtraction of values from control immunoprecipitations with only beads. Bars and error bars are averages and s.d. from at least three experiments. P -values were computed using the Student’s t -test. * P

    Article Snippet: Full-length human RNaseH1 complementary DNA (cDNA) was purchased from Origene, C-terminally myc tagged and cloned into the retroviral vector pLHCX (Clontech).

    Techniques: Chromatin Immunoprecipitation, Dot Blot, Immunoprecipitation, Transfection

    RNaseH1 depletion leads to RPA activation at ALT telomeres. ( a ) HeLa and U2OS cells were transfected with the indicated siRNAs and 48 and 72 h later protein extracts were prepared. Western blot analysis was performed using antibodies against RNaseH1, RPA32 phosphorylated at Serine 33 (pSer33), total RPA32 and KAP1 (loading control). The asterisk indicates a cross-reacting band. Cells treated for 6 h with 5 mM hydroxyurea (HU) were used as controls for pSer33 activation. ( b ) SiRNA transfected cells were subjected to indirect immunofluorescence using antibodies against TRF2 (to visualize telomeres; red) and pSer33 (green). DNA was counterstained with DAPI (blue). Arrows point to examples of pSer33 foci co-localizing with TRF2 (TIFs). Scale bar, 9 μm. Cells treated for 6 h with 5 mM HU were used as controls for pSer33 activation.

    Journal: Nature Communications

    Article Title: RNaseH1 regulates TERRA-telomeric DNA hybrids and telomere maintenance in ALT tumour cells

    doi: 10.1038/ncomms6220

    Figure Lengend Snippet: RNaseH1 depletion leads to RPA activation at ALT telomeres. ( a ) HeLa and U2OS cells were transfected with the indicated siRNAs and 48 and 72 h later protein extracts were prepared. Western blot analysis was performed using antibodies against RNaseH1, RPA32 phosphorylated at Serine 33 (pSer33), total RPA32 and KAP1 (loading control). The asterisk indicates a cross-reacting band. Cells treated for 6 h with 5 mM hydroxyurea (HU) were used as controls for pSer33 activation. ( b ) SiRNA transfected cells were subjected to indirect immunofluorescence using antibodies against TRF2 (to visualize telomeres; red) and pSer33 (green). DNA was counterstained with DAPI (blue). Arrows point to examples of pSer33 foci co-localizing with TRF2 (TIFs). Scale bar, 9 μm. Cells treated for 6 h with 5 mM HU were used as controls for pSer33 activation.

    Article Snippet: Full-length human RNaseH1 complementary DNA (cDNA) was purchased from Origene, C-terminally myc tagged and cloned into the retroviral vector pLHCX (Clontech).

    Techniques: Recombinase Polymerase Amplification, Activation Assay, Transfection, Western Blot, Immunofluorescence

    In vivo association and domain mapping of p53 and EAF2 protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of DNA damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.

    Journal: Endocrinology

    Article Title: Combined Loss of EAF2 and p53 Induces Prostate Carcinogenesis in Male Mice

    doi: 10.1210/en.2017-00409

    Figure Lengend Snippet: In vivo association and domain mapping of p53 and EAF2 protein interactions. (A) Coimmunoprecipitation of p53 with EAF2 in C4-2 cells treated with sip53 in C4-2 cells in the presence and absence of DNA damage induced by 1.0 μg/mL doxorubicin (Dx) for 6 hours. NEs were immunoprecipitated with immobilized anti-p53. Proteins were immunoblotted with anti-p53 and anti-EAF2. Lamin A/C served as loading control of nuclear proteins. (B) Domain mapping of the interaction between EAF2 and p53. GFP-p53 was cotransfected with Myc-tagged EAF2 deletion mutants (amino acids 113 to 260, 1 to 161, 161 to 260, and 1 to 245) into HEK293 cells. Myc-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. (C) Myc-EAF2 was cotransfected with GFP-tagged p53 deletion mutants [1 to 100, transcription activation domain (TAD); 100 to 292, DBD; and 292 to 393, C-terminal domain (CT)] into HEK293 cells. GFP-vec was added as a control plasmid to equalize the amount of transfection reagents for all cells to 2.5 μg total. Whole-cell lysates (WCL) were immunoprecipitated with anti-MYC or anti-GFP–conjugated agarose beads, and blots were probed with (B) anti-Myc or (C) anti-GFP antibody.

    Article Snippet: Full-length complementary DNA of human EAF2 and human p53 were subcloned into pCMV-Myc or pEGFP-C1 (Clontech, Mountain View, CA) vector to generate p-CMV-Myc-EAF2 and pEGFP-C1-p53 as previously described ( ).

    Techniques: In Vivo, Immunoprecipitation, Plasmid Preparation, Transfection, Activation Assay

    Schematic diagram of the construction of RGS14 -KO mice using the CRISPR-Cas9 method and identification of RGS14 expression. a One sgRNA targeting a region downstream of the 3′ end of exon 3 in the RGS14 mouse gene was designed and constructed. b After microinjection, a T7E1 assay indicated that four out of six pups contained cleavage products, suggesting a mixture of mutant and wild-type DNA templates in these mice. c Following subcloning of the PCR products, eight subclones of each mouse were sequenced. All of the subclones carried a single mutant allele, whereas two indels (#5–5, #5–6) produced frameshift mutations. Founder #5–5 was mated to a C57BL/6J mouse to obtain the F1 generation. d Gene sequencing for RGS14 expression levels in hearts from wild-type and RGS14 knockout groups. e Representative western blots for RGS14 expression levels in hearts from the RGS14 +/+ and RGS14 −/− groups

    Journal: Basic Research in Cardiology

    Article Title: Regulator of G protein signalling 14 attenuates cardiac remodelling through the MEK–ERK1/2 signalling pathway

    doi: 10.1007/s00395-016-0566-1

    Figure Lengend Snippet: Schematic diagram of the construction of RGS14 -KO mice using the CRISPR-Cas9 method and identification of RGS14 expression. a One sgRNA targeting a region downstream of the 3′ end of exon 3 in the RGS14 mouse gene was designed and constructed. b After microinjection, a T7E1 assay indicated that four out of six pups contained cleavage products, suggesting a mixture of mutant and wild-type DNA templates in these mice. c Following subcloning of the PCR products, eight subclones of each mouse were sequenced. All of the subclones carried a single mutant allele, whereas two indels (#5–5, #5–6) produced frameshift mutations. Founder #5–5 was mated to a C57BL/6J mouse to obtain the F1 generation. d Gene sequencing for RGS14 expression levels in hearts from wild-type and RGS14 knockout groups. e Representative western blots for RGS14 expression levels in hearts from the RGS14 +/+ and RGS14 −/− groups

    Article Snippet: Cardiac-specific RGS14 -overexpressing mice Full-length mouse RGS14 Complementary DNA (cDNA) (OriGene, MC204443) was ligated into the chicken β-actin gene (CAG) promoter expression vector, which was linearized and purified using the QIAquick Gel Extraction Kit (Qiagen, 28704).

    Techniques: Mouse Assay, CRISPR, Expressing, Construct, Mutagenesis, Subcloning, Polymerase Chain Reaction, Produced, Sequencing, Knock-Out, Western Blot

    Dual targeting of DNA topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. glauca (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA

    Journal: Plant Physiology

    Article Title: Acquisition, Conservation, and Loss of Dual-Targeted Proteins in Land Plants 1Acquisition, Conservation, and Loss of Dual-Targeted Proteins in Land Plants 1 [W]Acquisition, Conservation, and Loss of Dual-Targeted Proteins in Land Plants 1 [W] [OA]

    doi: 10.1104/pp.112.210997

    Figure Lengend Snippet: Dual targeting of DNA topoisomerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA topoisomerases (Top) from Arabidopsis (At), rice (Os), P. patens (Pp), P. glauca (Pg), C. reinhardtii (Cr), and C. variabilis (Cv) using MEGA

    Article Snippet: The translational start sites for all P. glauca genes were confirmed by 5′ RACE using CapFishing full-length complementary DNA ( ) premix kit (Seegene).

    Techniques:

    Dual targeting of DNA polymerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA polymerases (Pol) from Arabidopsis (At), rice (Os), P. patens (Pp), P. glauca (Pg), and C. reinhardtii (Cr) using MEGA 5 (see “Materials

    Journal: Plant Physiology

    Article Title: Acquisition, Conservation, and Loss of Dual-Targeted Proteins in Land Plants 1Acquisition, Conservation, and Loss of Dual-Targeted Proteins in Land Plants 1 [W]Acquisition, Conservation, and Loss of Dual-Targeted Proteins in Land Plants 1 [W] [OA]

    doi: 10.1104/pp.112.210997

    Figure Lengend Snippet: Dual targeting of DNA polymerase to mitochondria and plastids. A, Phylogenetic analysis of genes encoding DNA polymerases (Pol) from Arabidopsis (At), rice (Os), P. patens (Pp), P. glauca (Pg), and C. reinhardtii (Cr) using MEGA 5 (see “Materials

    Article Snippet: The translational start sites for all P. glauca genes were confirmed by 5′ RACE using CapFishing full-length complementary DNA ( ) premix kit (Seegene).

    Techniques: