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Becton Dickinson advantage gc rich kit
Advantage Gc Rich Kit, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Snippet: A standard solution was prepared using the Clontech Advantage® GC rich kit (BD-Biosciences Clontech, Palo Alto, CA).

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    Becton Dickinson advantage gc pcr kit
    <t>NF-IL6β</t> plays a role in transcription of cox-2 gene. ( A ) Overexpression of NF-IL6β enhances the cox-2 reporter activity. A431 cells were transfected with 0.2 µg of reporter vector pXC80 carrying cox-2 promoter (−80/+49 bp) together with expression vector of NF-IL6β in 1 ml of Opti-MEM medium. After medium change, cells were treated with or without EGF for 13 h. Cell lysates were then prepared, and luciferase activity was assayed. ( B ) Silencing of NF-IL6β expression attenuates EGF-induced cox-2 reporter activity. Cells were transfected with 1 µg of pXC918 carrying cox-2 promoter (−918/+49 bp) together with 1 µg of pSi-C or pSi-I. The upper panel shows that the transfection of NF-IL6β expression vectors attenuated the pXC918 reporter activity. The lower panel shows that the transcriptional products of cox-2 and NF-IL6β genes which were examined by <t>RT–PCR</t> analysis. The pSi-C represents the p Silencer ™ negative control plasmid. The pSi-I indicates the specific NF-IL6β knockdown expression vector. ( C ) Reduction of NF-IL6β expression decrease COX-2 expression. The transfection of pcDNA3/HA-NF-IL6β with pSi-C or pSi-I was performed in HeLa cells. Cellular lysates were harvested and analysed by western blotting probed with α-HA, α-COX-2 and α-β-actin antibodies. β-Actin was used as an internal loading control. ( D ) Silencing of c-Jun expression attenuates NF-IL6β-induced cox-2 reporter activity. A431 cells were transfected with 0.2 µg of pXC80, together with 0.5 µg of expression vector pSUPERc-Jun siRNA and 0.2 µg of each expression vector of c-Jun and NF-IL6β in 1 ml of Opti-MEM medium. Statistical significance between pSUPERc-Jun siRNA-transfected and untransfected cells was analysed by Student's t -test. ( E ) Cooperation of NF-IL6β with c-Jun in promoter activation of cox-2 gene. A431 cells were transfected with 0.2 µg of pXC80 reporter vector together with 0.05 µg of expression vector of NF/IL6β and 0.02 µg of expression vector of c-Jun in 1 ml of Opti-MEM medium. Cell lysates were then prepared, and luciferase activity was assayed. Statistical significance between c-Jun-transfected and untransfected cells was analysed by Student's t -test.
    Advantage Gc Pcr Kit, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/advantage gc pcr kit/product/Becton Dickinson
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    advantage gc pcr kit - by Bioz Stars, 2020-07
    85/100 stars
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    NF-IL6β plays a role in transcription of cox-2 gene. ( A ) Overexpression of NF-IL6β enhances the cox-2 reporter activity. A431 cells were transfected with 0.2 µg of reporter vector pXC80 carrying cox-2 promoter (−80/+49 bp) together with expression vector of NF-IL6β in 1 ml of Opti-MEM medium. After medium change, cells were treated with or without EGF for 13 h. Cell lysates were then prepared, and luciferase activity was assayed. ( B ) Silencing of NF-IL6β expression attenuates EGF-induced cox-2 reporter activity. Cells were transfected with 1 µg of pXC918 carrying cox-2 promoter (−918/+49 bp) together with 1 µg of pSi-C or pSi-I. The upper panel shows that the transfection of NF-IL6β expression vectors attenuated the pXC918 reporter activity. The lower panel shows that the transcriptional products of cox-2 and NF-IL6β genes which were examined by RT–PCR analysis. The pSi-C represents the p Silencer ™ negative control plasmid. The pSi-I indicates the specific NF-IL6β knockdown expression vector. ( C ) Reduction of NF-IL6β expression decrease COX-2 expression. The transfection of pcDNA3/HA-NF-IL6β with pSi-C or pSi-I was performed in HeLa cells. Cellular lysates were harvested and analysed by western blotting probed with α-HA, α-COX-2 and α-β-actin antibodies. β-Actin was used as an internal loading control. ( D ) Silencing of c-Jun expression attenuates NF-IL6β-induced cox-2 reporter activity. A431 cells were transfected with 0.2 µg of pXC80, together with 0.5 µg of expression vector pSUPERc-Jun siRNA and 0.2 µg of each expression vector of c-Jun and NF-IL6β in 1 ml of Opti-MEM medium. Statistical significance between pSUPERc-Jun siRNA-transfected and untransfected cells was analysed by Student's t -test. ( E ) Cooperation of NF-IL6β with c-Jun in promoter activation of cox-2 gene. A431 cells were transfected with 0.2 µg of pXC80 reporter vector together with 0.05 µg of expression vector of NF/IL6β and 0.02 µg of expression vector of c-Jun in 1 ml of Opti-MEM medium. Cell lysates were then prepared, and luciferase activity was assayed. Statistical significance between c-Jun-transfected and untransfected cells was analysed by Student's t -test.

    Journal: Nucleic Acids Research

    Article Title: Functional role of NF-IL6? and its sumoylation and acetylation modifications in promoter activation of cyclooxygenase 2 gene

    doi: 10.1093/nar/gkj422

    Figure Lengend Snippet: NF-IL6β plays a role in transcription of cox-2 gene. ( A ) Overexpression of NF-IL6β enhances the cox-2 reporter activity. A431 cells were transfected with 0.2 µg of reporter vector pXC80 carrying cox-2 promoter (−80/+49 bp) together with expression vector of NF-IL6β in 1 ml of Opti-MEM medium. After medium change, cells were treated with or without EGF for 13 h. Cell lysates were then prepared, and luciferase activity was assayed. ( B ) Silencing of NF-IL6β expression attenuates EGF-induced cox-2 reporter activity. Cells were transfected with 1 µg of pXC918 carrying cox-2 promoter (−918/+49 bp) together with 1 µg of pSi-C or pSi-I. The upper panel shows that the transfection of NF-IL6β expression vectors attenuated the pXC918 reporter activity. The lower panel shows that the transcriptional products of cox-2 and NF-IL6β genes which were examined by RT–PCR analysis. The pSi-C represents the p Silencer ™ negative control plasmid. The pSi-I indicates the specific NF-IL6β knockdown expression vector. ( C ) Reduction of NF-IL6β expression decrease COX-2 expression. The transfection of pcDNA3/HA-NF-IL6β with pSi-C or pSi-I was performed in HeLa cells. Cellular lysates were harvested and analysed by western blotting probed with α-HA, α-COX-2 and α-β-actin antibodies. β-Actin was used as an internal loading control. ( D ) Silencing of c-Jun expression attenuates NF-IL6β-induced cox-2 reporter activity. A431 cells were transfected with 0.2 µg of pXC80, together with 0.5 µg of expression vector pSUPERc-Jun siRNA and 0.2 µg of each expression vector of c-Jun and NF-IL6β in 1 ml of Opti-MEM medium. Statistical significance between pSUPERc-Jun siRNA-transfected and untransfected cells was analysed by Student's t -test. ( E ) Cooperation of NF-IL6β with c-Jun in promoter activation of cox-2 gene. A431 cells were transfected with 0.2 µg of pXC80 reporter vector together with 0.05 µg of expression vector of NF/IL6β and 0.02 µg of expression vector of c-Jun in 1 ml of Opti-MEM medium. Cell lysates were then prepared, and luciferase activity was assayed. Statistical significance between c-Jun-transfected and untransfected cells was analysed by Student's t -test.

    Article Snippet: NF-IL6β was generated from human liver cDNA library by PCR using BD Advantage GC™ PCR kit and using the following oligonucleotides: 5′-CGGGATCCAGCGCCGCGCTCTTCAGCCTG-3′ and 5′-GGCCTCGAGGCCGCGCGTTACCGGCAGTC-3′.

    Techniques: Over Expression, Activity Assay, Transfection, Plasmid Preparation, Expressing, Luciferase, Reverse Transcription Polymerase Chain Reaction, Negative Control, Western Blot, Activation Assay

    NF-IL6β binds to the cox-2 gene promoter in vivo . ( A ) ChIP analysis was performed as described in Materials and Methods. The upper panel indicates the scheme of 5′-flanking region of cox-2 gene, and the location of designed primers for PCR. Chromatin from A431 cells with or without EGF treatment was immunoprecipitated with specific antibodies as indicated and the cox-2 promoter region was amplified by PCR. ( B ) Purified p300 proteins cannot interact with suNF-IL6β. The pull-down assay was performed using the mixtures of purified p300 protein incubated with in vitro -translated HA/NF-IL6β with or without sumoylation enzymes as described in Materials and Methods. The ‘C’ represents the control products of in vitro transcription/translation reaction with pCDNA3/HA vector; the ‘Su-C’ represents the products were performed by in vitro -translated control products in in vitro -sumoylation reaction. Western blots of reaction mixture (left panel) and α-p300-immunoprecipitated pellet (right panel), using α-HA antibodies, are indicated.

    Journal: Nucleic Acids Research

    Article Title: Functional role of NF-IL6? and its sumoylation and acetylation modifications in promoter activation of cyclooxygenase 2 gene

    doi: 10.1093/nar/gkj422

    Figure Lengend Snippet: NF-IL6β binds to the cox-2 gene promoter in vivo . ( A ) ChIP analysis was performed as described in Materials and Methods. The upper panel indicates the scheme of 5′-flanking region of cox-2 gene, and the location of designed primers for PCR. Chromatin from A431 cells with or without EGF treatment was immunoprecipitated with specific antibodies as indicated and the cox-2 promoter region was amplified by PCR. ( B ) Purified p300 proteins cannot interact with suNF-IL6β. The pull-down assay was performed using the mixtures of purified p300 protein incubated with in vitro -translated HA/NF-IL6β with or without sumoylation enzymes as described in Materials and Methods. The ‘C’ represents the control products of in vitro transcription/translation reaction with pCDNA3/HA vector; the ‘Su-C’ represents the products were performed by in vitro -translated control products in in vitro -sumoylation reaction. Western blots of reaction mixture (left panel) and α-p300-immunoprecipitated pellet (right panel), using α-HA antibodies, are indicated.

    Article Snippet: NF-IL6β was generated from human liver cDNA library by PCR using BD Advantage GC™ PCR kit and using the following oligonucleotides: 5′-CGGGATCCAGCGCCGCGCTCTTCAGCCTG-3′ and 5′-GGCCTCGAGGCCGCGCGTTACCGGCAGTC-3′.

    Techniques: In Vivo, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Immunoprecipitation, Amplification, Purification, Pull Down Assay, Incubation, In Vitro, Plasmid Preparation, Western Blot

    ( A ) EGF-induced COX-2 expression and C/EBP protein levels in A431 cells. Cell lysates were harvested after EGF treatment. Antibodies to recognize the COX-1, COX-2, c-Jun and C/EBPs protein levels performed by western blot analysis. ( B ) SB203580, p38 inhibitor, attenuates EGF-induced cox-2 transcription in a dose-dependent manner. A431 cells deprived of serum were pretreated with p38 inhibitor SB203580 for 30 min prior to stimulation with EGF for 90 min. After stimulation, cells were lysed, and total RNA was prepared for RT–PCR analysis of NF-IL6β and cox-2 mRNA levels.

    Journal: Nucleic Acids Research

    Article Title: Functional role of NF-IL6? and its sumoylation and acetylation modifications in promoter activation of cyclooxygenase 2 gene

    doi: 10.1093/nar/gkj422

    Figure Lengend Snippet: ( A ) EGF-induced COX-2 expression and C/EBP protein levels in A431 cells. Cell lysates were harvested after EGF treatment. Antibodies to recognize the COX-1, COX-2, c-Jun and C/EBPs protein levels performed by western blot analysis. ( B ) SB203580, p38 inhibitor, attenuates EGF-induced cox-2 transcription in a dose-dependent manner. A431 cells deprived of serum were pretreated with p38 inhibitor SB203580 for 30 min prior to stimulation with EGF for 90 min. After stimulation, cells were lysed, and total RNA was prepared for RT–PCR analysis of NF-IL6β and cox-2 mRNA levels.

    Article Snippet: NF-IL6β was generated from human liver cDNA library by PCR using BD Advantage GC™ PCR kit and using the following oligonucleotides: 5′-CGGGATCCAGCGCCGCGCTCTTCAGCCTG-3′ and 5′-GGCCTCGAGGCCGCGCGTTACCGGCAGTC-3′.

    Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction