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sp-6 rna polymerase reagent kits for synthesis of labelled and unlabelled rnas  (Promega)

 
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    Promega sp-6 rna polymerase reagent kits for synthesis of labelled and unlabelled rnas
    CD437 regulation of chimeric rabbit β-globin-p21 WAF1/CIP1 mRNA . Two independent sublines derived from transfection of RGB-WAF (581-2004) plasmid [31.4(A4) and 31.4(C5)] or RBG plasmid [29.6(1) and 29.6(2)] were grown either in the absence (-) or presence (+) of 1 μM CD437 for 48 hours. Total <t>RNAs</t> were prepared and expression of RBG transcripts analyzed by northern blot hybridization as described . Levels <t>of</t> <t>RNA</t> loading in each lane were assessed by signals from hybridization with ribosomal phosphoprotein 36B4 cDNA
    Sp 6 Rna Polymerase Reagent Kits For Synthesis Of Labelled And Unlabelled Rnas, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sp-6+rna+polymerase/sp+6+rna+polymerase+reagent+kits+for+synthesis+of+labelled+and+unlabelled+rnas/pmc02927583-159-18-23
    Average 90 stars, based on 1 article reviews
    sp-6 rna polymerase reagent kits for synthesis of labelled and unlabelled rnas - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "Novel cis-trans interactions are involved in post-transcriptional regulation of cyclin-dependent kinase inhibitor p21 WAF1/CIP1 mRNA"

    Article Title: Novel cis-trans interactions are involved in post-transcriptional regulation of cyclin-dependent kinase inhibitor p21 WAF1/CIP1 mRNA

    Journal: Journal of Molecular Signaling

    doi: 10.1186/1750-2187-5-12

    CD437 regulation of chimeric rabbit β-globin-p21 WAF1/CIP1 mRNA . Two independent sublines derived from transfection of RGB-WAF (581-2004) plasmid [31.4(A4) and 31.4(C5)] or RBG plasmid [29.6(1) and 29.6(2)] were grown either in the absence (-) or presence (+) of 1 μM CD437 for 48 hours. Total RNAs were prepared and expression of RBG transcripts analyzed by northern blot hybridization as described . Levels of RNA loading in each lane were assessed by signals from hybridization with ribosomal phosphoprotein 36B4 cDNA
    Figure Legend Snippet: CD437 regulation of chimeric rabbit β-globin-p21 WAF1/CIP1 mRNA . Two independent sublines derived from transfection of RGB-WAF (581-2004) plasmid [31.4(A4) and 31.4(C5)] or RBG plasmid [29.6(1) and 29.6(2)] were grown either in the absence (-) or presence (+) of 1 μM CD437 for 48 hours. Total RNAs were prepared and expression of RBG transcripts analyzed by northern blot hybridization as described . Levels of RNA loading in each lane were assessed by signals from hybridization with ribosomal phosphoprotein 36B4 cDNA

    Techniques Used: Derivative Assay, Transfection, Plasmid Preparation, Expressing, Northern Blot, Hybridization

    Multiple cell cytoplasmic proteins bind to radiolabelled p21 WAF1/CIP1 3'-UTR and GADD45 5'-UTR sense strand riboprobes . Transcription and labeling of the indicated probes as well as the in vitro binding reactions were as described in the Methods. The probes utilized and treatment times with CD437 (hr) are indicated above each lane in the respective panels. The cytoplasmic protein extracts were prepared from untreated (lanes marked 0 hours) or CD437-treated MDA-MB-468 (panels A-C), MCF-7 (panel C) and HL-60R (panel C) cells. Panel A, the protein extracts in lanes 4-6 and11-13 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Probe 14.2 RNA is loaded in lane 14. Panel B , the protein extracts in lanes 3 and 6 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Panel C , lanes 6, 12 represent the labeled probe only. The approximate migration of molecular weight standards is marked on the left side of respective panel, while the * and ** on the right side of each panel denote locations of the putative 85 kD and 55 kD complexes, respectively.
    Figure Legend Snippet: Multiple cell cytoplasmic proteins bind to radiolabelled p21 WAF1/CIP1 3'-UTR and GADD45 5'-UTR sense strand riboprobes . Transcription and labeling of the indicated probes as well as the in vitro binding reactions were as described in the Methods. The probes utilized and treatment times with CD437 (hr) are indicated above each lane in the respective panels. The cytoplasmic protein extracts were prepared from untreated (lanes marked 0 hours) or CD437-treated MDA-MB-468 (panels A-C), MCF-7 (panel C) and HL-60R (panel C) cells. Panel A, the protein extracts in lanes 4-6 and11-13 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Probe 14.2 RNA is loaded in lane 14. Panel B , the protein extracts in lanes 3 and 6 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Panel C , lanes 6, 12 represent the labeled probe only. The approximate migration of molecular weight standards is marked on the left side of respective panel, while the * and ** on the right side of each panel denote locations of the putative 85 kD and 55 kD complexes, respectively.

    Techniques Used: Labeling, In Vitro, Binding Assay, Incubation, Migration, Molecular Weight

    Related Articles

    In Vitro:

    Article Title: The Lipid Droplet Binding Domain of Hepatitis C Virus Core Protein Is a Major Determinant for Efficient Virus Assembly
    Article Snippet: In vitro transcription reaction mixtures with SFV plasmids contained 1 unit of 5 SP-6 buffer (Promega), 3.125 mM of each nucleotide, 1 unit of RNasin (Promega, Mannheim, Germany) per l, 1 mM cap analogue m7G(5 )ppp(5 )G (Roche Applied Science) per l, 0.1 g of plasmid DNA/ l, and 1.8 units of SP-6 RNA polymerase (Promega) per l. After 2 h at 37 °C, an additional 1 unit of SP-6 RNA polymerase/ l was added, and the reaction mixture was incubated for another 2 h. Transcription was terminated by a 30-min incubation at 37 °C with 1.2 units of RNase-free DNase (Promega) per g of plasmid DNA.

    Article Title: Activation of the cytochrome c gene by electrical stimulation in neonatal rat cardiac myocytes. Role of NRF-1 and c-Jun.
    Article Snippet: Radiolabeled RNA transcripts were synthesized from template DNA linearized with Eco0109 as described by the Promega technical manual using Sp-6 RNA polymerase, ATP, GTP, UTP and [a-32P]CTP (0.5 mM each) in the presence of RNasin and spermidine.

    Plasmid Preparation:

    Article Title: The Lipid Droplet Binding Domain of Hepatitis C Virus Core Protein Is a Major Determinant for Efficient Virus Assembly
    Article Snippet: In vitro transcription reaction mixtures with SFV plasmids contained 1 unit of 5 SP-6 buffer (Promega), 3.125 mM of each nucleotide, 1 unit of RNasin (Promega, Mannheim, Germany) per l, 1 mM cap analogue m7G(5 )ppp(5 )G (Roche Applied Science) per l, 0.1 g of plasmid DNA/ l, and 1.8 units of SP-6 RNA polymerase (Promega) per l. After 2 h at 37 °C, an additional 1 unit of SP-6 RNA polymerase/ l was added, and the reaction mixture was incubated for another 2 h. Transcription was terminated by a 30-min incubation at 37 °C with 1.2 units of RNase-free DNase (Promega) per g of plasmid DNA.

    Article Title: Activation of the cytochrome c gene by electrical stimulation in neonatal rat cardiac myocytes. Role of NRF-1 and c-Jun.
    Article Snippet: Radiolabeled RNA transcripts were synthesized from template DNA linearized with Eco0109 as described by the Promega technical manual using Sp-6 RNA polymerase, ATP, GTP, UTP and [a-32P]CTP (0.5 mM each) in the presence of RNasin and spermidine.

    Incubation:

    Article Title: The Lipid Droplet Binding Domain of Hepatitis C Virus Core Protein Is a Major Determinant for Efficient Virus Assembly
    Article Snippet: In vitro transcription reaction mixtures with SFV plasmids contained 1 unit of 5 SP-6 buffer (Promega), 3.125 mM of each nucleotide, 1 unit of RNasin (Promega, Mannheim, Germany) per l, 1 mM cap analogue m7G(5 )ppp(5 )G (Roche Applied Science) per l, 0.1 g of plasmid DNA/ l, and 1.8 units of SP-6 RNA polymerase (Promega) per l. After 2 h at 37 °C, an additional 1 unit of SP-6 RNA polymerase/ l was added, and the reaction mixture was incubated for another 2 h. Transcription was terminated by a 30-min incubation at 37 °C with 1.2 units of RNase-free DNase (Promega) per g of plasmid DNA.

    Article Title: Activation of the cytochrome c gene by electrical stimulation in neonatal rat cardiac myocytes. Role of NRF-1 and c-Jun.
    Article Snippet: Radiolabeled RNA transcripts were synthesized from template DNA linearized with Eco0109 as described by the Promega technical manual using Sp-6 RNA polymerase, ATP, GTP, UTP and [a-32P]CTP (0.5 mM each) in the presence of RNasin and spermidine.



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    Image Search Results


    CD437 regulation of chimeric rabbit β-globin-p21 WAF1/CIP1 mRNA . Two independent sublines derived from transfection of RGB-WAF (581-2004) plasmid [31.4(A4) and 31.4(C5)] or RBG plasmid [29.6(1) and 29.6(2)] were grown either in the absence (-) or presence (+) of 1 μM CD437 for 48 hours. Total RNAs were prepared and expression of RBG transcripts analyzed by northern blot hybridization as described . Levels of RNA loading in each lane were assessed by signals from hybridization with ribosomal phosphoprotein 36B4 cDNA

    Journal: Journal of Molecular Signaling

    Article Title: Novel cis-trans interactions are involved in post-transcriptional regulation of cyclin-dependent kinase inhibitor p21 WAF1/CIP1 mRNA

    doi: 10.1186/1750-2187-5-12

    Figure Lengend Snippet: CD437 regulation of chimeric rabbit β-globin-p21 WAF1/CIP1 mRNA . Two independent sublines derived from transfection of RGB-WAF (581-2004) plasmid [31.4(A4) and 31.4(C5)] or RBG plasmid [29.6(1) and 29.6(2)] were grown either in the absence (-) or presence (+) of 1 μM CD437 for 48 hours. Total RNAs were prepared and expression of RBG transcripts analyzed by northern blot hybridization as described . Levels of RNA loading in each lane were assessed by signals from hybridization with ribosomal phosphoprotein 36B4 cDNA

    Article Snippet: The T 7 , T 3 , and SP-6 RNA polymerase reagent kits for synthesis of labelled and unlabelled RNAs were purchased from Promega (Madison, WI).

    Techniques: Derivative Assay, Transfection, Plasmid Preparation, Expressing, Northern Blot, Hybridization

    Multiple cell cytoplasmic proteins bind to radiolabelled p21 WAF1/CIP1 3'-UTR and GADD45 5'-UTR sense strand riboprobes . Transcription and labeling of the indicated probes as well as the in vitro binding reactions were as described in the Methods. The probes utilized and treatment times with CD437 (hr) are indicated above each lane in the respective panels. The cytoplasmic protein extracts were prepared from untreated (lanes marked 0 hours) or CD437-treated MDA-MB-468 (panels A-C), MCF-7 (panel C) and HL-60R (panel C) cells. Panel A, the protein extracts in lanes 4-6 and11-13 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Probe 14.2 RNA is loaded in lane 14. Panel B , the protein extracts in lanes 3 and 6 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Panel C , lanes 6, 12 represent the labeled probe only. The approximate migration of molecular weight standards is marked on the left side of respective panel, while the * and ** on the right side of each panel denote locations of the putative 85 kD and 55 kD complexes, respectively.

    Journal: Journal of Molecular Signaling

    Article Title: Novel cis-trans interactions are involved in post-transcriptional regulation of cyclin-dependent kinase inhibitor p21 WAF1/CIP1 mRNA

    doi: 10.1186/1750-2187-5-12

    Figure Lengend Snippet: Multiple cell cytoplasmic proteins bind to radiolabelled p21 WAF1/CIP1 3'-UTR and GADD45 5'-UTR sense strand riboprobes . Transcription and labeling of the indicated probes as well as the in vitro binding reactions were as described in the Methods. The probes utilized and treatment times with CD437 (hr) are indicated above each lane in the respective panels. The cytoplasmic protein extracts were prepared from untreated (lanes marked 0 hours) or CD437-treated MDA-MB-468 (panels A-C), MCF-7 (panel C) and HL-60R (panel C) cells. Panel A, the protein extracts in lanes 4-6 and11-13 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Probe 14.2 RNA is loaded in lane 14. Panel B , the protein extracts in lanes 3 and 6 were pre-incubated with 200-fold excess of the respective unlabeled probe RNAs, while the protein extracts in lane 7 were pre-incubated with 200-fold excess of the unlabeled 3'-UTR of c-myc RNA followed by incubation with the indicated labeled probe RNAs. Panel C , lanes 6, 12 represent the labeled probe only. The approximate migration of molecular weight standards is marked on the left side of respective panel, while the * and ** on the right side of each panel denote locations of the putative 85 kD and 55 kD complexes, respectively.

    Article Snippet: The T 7 , T 3 , and SP-6 RNA polymerase reagent kits for synthesis of labelled and unlabelled RNAs were purchased from Promega (Madison, WI).

    Techniques: Labeling, In Vitro, Binding Assay, Incubation, Migration, Molecular Weight