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TaKaRa human mrna
Human Mrna, supplied by TaKaRa, 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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human mrna - by Bioz Stars, 2020-09
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Clone Assay:

Article Title: Photocleavage Based Affinity Purification and Printing of Cell-free Expressed Proteins: Application to Proteome Microarrays
Article Snippet: .. Source DNA for human gene cloning was either human mRNA from Clontech Laboratories, Inc. (Mountain View, CA) or cDNAs from the I.M.A.G.E. .. The Ultrafree-MC, Durapore PVDF membrane, 500 µL capacity, micro-centrifuge filtration devices were from Millipore (Billerica, MA).

Northern Blot:

Article Title: Human Ligands of the Notch Receptor
Article Snippet: .. To analyze the expression of these ligands in human tissues, we probed Northern blots containing adult human mRNA from several tissues (Clontech). ..

Article Title: The Orphan Nuclear Receptor Ear-2 Is a Negative Coregulator for Thyroid Hormone Nuclear Receptor Function
Article Snippet: .. A human poly(A) RNA-blotted membrane, MTN Blots (Clontech), was used for Northern blot analysis. .. The membrane was hybridized with human Ear-2 or TRβ1 cDNA labeled with [32 P]dCTP by random priming and was subsequently washed with 2× SSC (1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate)–0.05% sodium dodecyl sulfate (SDS) at room temperature for 30 min and then 0.1× SSC–0.1% SDS at 50°C for 30 min.

Expressing:

Article Title: Human Ligands of the Notch Receptor
Article Snippet: .. To analyze the expression of these ligands in human tissues, we probed Northern blots containing adult human mRNA from several tissues (Clontech). ..

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    TaKaRa human normal thymus polyadenylated mrna species
    Expression pattern of the human H430 retropseudogene. (A) Correspondence of the DNA and RNA probes with the human H430 sequences. (B) An RNase protection analysis of H430 transcripts was performed with the ES165 riboprobe and 7 μg of the indicated <t>polyadenylated</t> RNA. The sizes of the specifically protected fragments are in nucleotides. (C and D) Northern blot analyses of H430 <t>mRNA</t> species. Polyadenylated RNA samples from the indicated human cells lines (5 μg per lane) and normal tissues (2 μg per lane) were hybridized with the 32 P-labeled ES165 DNA probe. Transcripts detected in the different samples are indicated by arrows. Sizes are in kilobases. Human GAPDH and β-actin were used as the internal controls to normalize for variations in RNA amounts.
    Human Normal Thymus Polyadenylated Mrna Species, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 80 stars, based on 1 article reviews
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    85
    TaKaRa 12 lane human poly a rna filter
    Example of transcripts (ncINT1Ms2) identified by the DMD-GEx array. A) a series of consecutive probes in the genome with fluorescence intensities that rank above the 90th percentile over all probes on the array (indicated with a dashed red line) and mapping within intron 1 M (chrX:33057364-33059742). B) location of the transcripts identified from poly A+ <t>RNA</t> with respect to the DMD gene isoforms. Sense transcripts are represented by blue bars, whereas antisense transcripts are indicated by green bars. The transcripts marked with an asterisk and a double cross were characterized by Northern blotting, and RACE PCR.
    12 Lane Human Poly A Rna Filter, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa mrna half life determinations human hela cells
    <t>mRNA–mRNA</t> duplexes create SBSs and trigger SMD a , Diagram of FLUC-SOWAHC 3' UTR mRNA harboring 12 copies of MS2bs base-paired with CDCP1 , TUFT1 or PPID mRNA via complementary 3' UTR Alu elements. b , Western blot (upper) or RT-sqPCR(lower) before (−) or after immunoprecipitation (IP) of lysates of formaldehyde-crosslinked <t>HeLa</t> cells that had been transiently transfected with pFlag-MS2-hMGFP and pFLUC-SOWAHC 3' UTR, pFLUC-SOWAHC 3' UTR-MS2bs or pFLUC-MS2bs. IP was performed using either anti-Flag or, as a control for nonspecific IP, mouse immunoglobulin G (mIgG). c , essentially as in b except pFLUC-SOWAHC 3' UTR-MS2bs or pFLUC-SOWAHC 3' UTR(ΔAlu)-MS2bs was used. d , essentially as in b except cells were transfected with Control or STAU1 siRNA. Error bars, s.e.m.; # of independently performed experiments = 3; P
    Mrna Half Life Determinations Human Hela Cells, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression pattern of the human H430 retropseudogene. (A) Correspondence of the DNA and RNA probes with the human H430 sequences. (B) An RNase protection analysis of H430 transcripts was performed with the ES165 riboprobe and 7 μg of the indicated polyadenylated RNA. The sizes of the specifically protected fragments are in nucleotides. (C and D) Northern blot analyses of H430 mRNA species. Polyadenylated RNA samples from the indicated human cells lines (5 μg per lane) and normal tissues (2 μg per lane) were hybridized with the 32 P-labeled ES165 DNA probe. Transcripts detected in the different samples are indicated by arrows. Sizes are in kilobases. Human GAPDH and β-actin were used as the internal controls to normalize for variations in RNA amounts.

    Journal: Molecular and Cellular Biology

    Article Title: Characterization of SRp46, a Novel Human SR Splicing Factor Encoded by a PR264/SC35 Retropseudogene

    doi:

    Figure Lengend Snippet: Expression pattern of the human H430 retropseudogene. (A) Correspondence of the DNA and RNA probes with the human H430 sequences. (B) An RNase protection analysis of H430 transcripts was performed with the ES165 riboprobe and 7 μg of the indicated polyadenylated RNA. The sizes of the specifically protected fragments are in nucleotides. (C and D) Northern blot analyses of H430 mRNA species. Polyadenylated RNA samples from the indicated human cells lines (5 μg per lane) and normal tissues (2 μg per lane) were hybridized with the 32 P-labeled ES165 DNA probe. Transcripts detected in the different samples are indicated by arrows. Sizes are in kilobases. Human GAPDH and β-actin were used as the internal controls to normalize for variations in RNA amounts.

    Article Snippet: Rapid amplification of cDNA ends (RACE)-PCR experiments were performed with human normal thymus polyadenylated mRNA species and the Marathon cDNA amplification kit (Clontech) according to the manufacturer’s instructions.

    Techniques: Expressing, Northern Blot, Labeling

    Example of transcripts (ncINT1Ms2) identified by the DMD-GEx array. A) a series of consecutive probes in the genome with fluorescence intensities that rank above the 90th percentile over all probes on the array (indicated with a dashed red line) and mapping within intron 1 M (chrX:33057364-33059742). B) location of the transcripts identified from poly A+ RNA with respect to the DMD gene isoforms. Sense transcripts are represented by blue bars, whereas antisense transcripts are indicated by green bars. The transcripts marked with an asterisk and a double cross were characterized by Northern blotting, and RACE PCR.

    Journal: PLoS ONE

    Article Title: The DMD Locus Harbours Multiple Long Non-Coding RNAs Which Orchestrate and Control Transcription of Muscle Dystrophin mRNA Isoforms

    doi: 10.1371/journal.pone.0045328

    Figure Lengend Snippet: Example of transcripts (ncINT1Ms2) identified by the DMD-GEx array. A) a series of consecutive probes in the genome with fluorescence intensities that rank above the 90th percentile over all probes on the array (indicated with a dashed red line) and mapping within intron 1 M (chrX:33057364-33059742). B) location of the transcripts identified from poly A+ RNA with respect to the DMD gene isoforms. Sense transcripts are represented by blue bars, whereas antisense transcripts are indicated by green bars. The transcripts marked with an asterisk and a double cross were characterized by Northern blotting, and RACE PCR.

    Article Snippet: Northern Blotting Sense transcripts originating from the intronic regions harbouring the DMD gene isoform promoters and antisense transcripts were validated by Northern blotting, using a 12-lane human poly A+ RNA filter (Clontech).

    Techniques: Fluorescence, Northern Blot, Polymerase Chain Reaction

    Northern blotting analyses on a 12-lane human poly A+ RNA filter using probes designed on ncRNAs originating near the first exons of DMD isoforms and antisense transcripts. Transcript ncINT1Ms2 is located near full-length DMD gene isoform Dp427p, whereas ncINT44s and ncINT44s2 surround isoform Dp140. Transcripts ncINT55s and ncINT55as are located upstream the Dp116 isoform. Nc3UTRas overlaps with 3′UTR in antisense direction with respect to the DMD gene. All transcripts were expressed in at least one tissue in which DMD isoforms were also expressed, but also in the liver, kidney, spleen and placenta. ncINT1Ms2, ncINT44s2, ncINT55s and nc3UTRas were found to be expressed in multiple isoforms, while one single isoform was detected for ncINT44s and ncINT55as.

    Journal: PLoS ONE

    Article Title: The DMD Locus Harbours Multiple Long Non-Coding RNAs Which Orchestrate and Control Transcription of Muscle Dystrophin mRNA Isoforms

    doi: 10.1371/journal.pone.0045328

    Figure Lengend Snippet: Northern blotting analyses on a 12-lane human poly A+ RNA filter using probes designed on ncRNAs originating near the first exons of DMD isoforms and antisense transcripts. Transcript ncINT1Ms2 is located near full-length DMD gene isoform Dp427p, whereas ncINT44s and ncINT44s2 surround isoform Dp140. Transcripts ncINT55s and ncINT55as are located upstream the Dp116 isoform. Nc3UTRas overlaps with 3′UTR in antisense direction with respect to the DMD gene. All transcripts were expressed in at least one tissue in which DMD isoforms were also expressed, but also in the liver, kidney, spleen and placenta. ncINT1Ms2, ncINT44s2, ncINT55s and nc3UTRas were found to be expressed in multiple isoforms, while one single isoform was detected for ncINT44s and ncINT55as.

    Article Snippet: Northern Blotting Sense transcripts originating from the intronic regions harbouring the DMD gene isoform promoters and antisense transcripts were validated by Northern blotting, using a 12-lane human poly A+ RNA filter (Clontech).

    Techniques: Northern Blot

    mRNA–mRNA duplexes create SBSs and trigger SMD a , Diagram of FLUC-SOWAHC 3' UTR mRNA harboring 12 copies of MS2bs base-paired with CDCP1 , TUFT1 or PPID mRNA via complementary 3' UTR Alu elements. b , Western blot (upper) or RT-sqPCR(lower) before (−) or after immunoprecipitation (IP) of lysates of formaldehyde-crosslinked HeLa cells that had been transiently transfected with pFlag-MS2-hMGFP and pFLUC-SOWAHC 3' UTR, pFLUC-SOWAHC 3' UTR-MS2bs or pFLUC-MS2bs. IP was performed using either anti-Flag or, as a control for nonspecific IP, mouse immunoglobulin G (mIgG). c , essentially as in b except pFLUC-SOWAHC 3' UTR-MS2bs or pFLUC-SOWAHC 3' UTR(ΔAlu)-MS2bs was used. d , essentially as in b except cells were transfected with Control or STAU1 siRNA. Error bars, s.e.m.; # of independently performed experiments = 3; P

    Journal: Nature structural & molecular biology

    Article Title: mRNA-mRNA duplexes that auto-elicit Staufen1-mediated mRNA decay

    doi: 10.1038/nsmb.2664

    Figure Lengend Snippet: mRNA–mRNA duplexes create SBSs and trigger SMD a , Diagram of FLUC-SOWAHC 3' UTR mRNA harboring 12 copies of MS2bs base-paired with CDCP1 , TUFT1 or PPID mRNA via complementary 3' UTR Alu elements. b , Western blot (upper) or RT-sqPCR(lower) before (−) or after immunoprecipitation (IP) of lysates of formaldehyde-crosslinked HeLa cells that had been transiently transfected with pFlag-MS2-hMGFP and pFLUC-SOWAHC 3' UTR, pFLUC-SOWAHC 3' UTR-MS2bs or pFLUC-MS2bs. IP was performed using either anti-Flag or, as a control for nonspecific IP, mouse immunoglobulin G (mIgG). c , essentially as in b except pFLUC-SOWAHC 3' UTR-MS2bs or pFLUC-SOWAHC 3' UTR(ΔAlu)-MS2bs was used. d , essentially as in b except cells were transfected with Control or STAU1 siRNA. Error bars, s.e.m.; # of independently performed experiments = 3; P

    Article Snippet: Cell culture, transient transfections, formaldehyde crosslinking, and mRNA half-life determinations Human HeLa cells, HeLa Tet-off cells (Clontech), HeLa cells stably expressing IRE-Gl Norm mRNA , or HaCaT cells were grown in DMEM (Gibco-BRL) containing 10% fetal bovine serum (Gibco-BRL) at 37°C and in 5% CO2 .

    Techniques: Western Blot, Immunoprecipitation, Transfection

    SOWAHC mRNA reduces the abundance of mRNAs to which it is predicted to base-pair via partially complementary 3' UTR Alu elements a , Diagram of computationally predicted putative mRNA–mRNA duplexes that form in human cells via base-pairing between partially complementary 3' UTR Alu elements ( Supplementary Table S1 ). b , Histogram representation of RT-sqPCR analyses of the indicated mRNA using lysates of HeLa cells treated with the specified siRNA (see Supplementary Fig. 1o for data using alternative siRNAs). The level of each mRNA was normalized to the level of GAPDH mRNA, and normalized levels in the presence of Control siRNA are defined as 100. Black arrows denote results that implicate mRNA–mRNA duplex formation. c , Western blot (WB) using the designated antibody and lysates analyzed in c. Calnexin serves as a loading control. d , Histogram representation of RT-sqPCR analyses of lysates of HeLa cells that had been transiently transfected with the specified siRNA and pFLUC-SOWAHC 3' UTR, pFLUC-SOWAHC 3' UTR(ΔAlu) or, as a negative control, pFLUC-No SBS. The level of each mRNA was normalized to the level of MUP mRNA (from the phCMV-MUP reference plasmid), and normalized levels in the presence of Control siRNA + pFLUC-No SBS are defined as 100. Error bars, s.e.m.; # of independently performed experiments = 3; *, P

    Journal: Nature structural & molecular biology

    Article Title: mRNA-mRNA duplexes that auto-elicit Staufen1-mediated mRNA decay

    doi: 10.1038/nsmb.2664

    Figure Lengend Snippet: SOWAHC mRNA reduces the abundance of mRNAs to which it is predicted to base-pair via partially complementary 3' UTR Alu elements a , Diagram of computationally predicted putative mRNA–mRNA duplexes that form in human cells via base-pairing between partially complementary 3' UTR Alu elements ( Supplementary Table S1 ). b , Histogram representation of RT-sqPCR analyses of the indicated mRNA using lysates of HeLa cells treated with the specified siRNA (see Supplementary Fig. 1o for data using alternative siRNAs). The level of each mRNA was normalized to the level of GAPDH mRNA, and normalized levels in the presence of Control siRNA are defined as 100. Black arrows denote results that implicate mRNA–mRNA duplex formation. c , Western blot (WB) using the designated antibody and lysates analyzed in c. Calnexin serves as a loading control. d , Histogram representation of RT-sqPCR analyses of lysates of HeLa cells that had been transiently transfected with the specified siRNA and pFLUC-SOWAHC 3' UTR, pFLUC-SOWAHC 3' UTR(ΔAlu) or, as a negative control, pFLUC-No SBS. The level of each mRNA was normalized to the level of MUP mRNA (from the phCMV-MUP reference plasmid), and normalized levels in the presence of Control siRNA + pFLUC-No SBS are defined as 100. Error bars, s.e.m.; # of independently performed experiments = 3; *, P

    Article Snippet: Cell culture, transient transfections, formaldehyde crosslinking, and mRNA half-life determinations Human HeLa cells, HeLa Tet-off cells (Clontech), HeLa cells stably expressing IRE-Gl Norm mRNA , or HaCaT cells were grown in DMEM (Gibco-BRL) containing 10% fetal bovine serum (Gibco-BRL) at 37°C and in 5% CO2 .

    Techniques: Western Blot, Transfection, Negative Control, Plasmid Preparation

    mRNA–mRNA duplexes create SBSs and trigger SMD a , Western blot (upper) or RT-sqPCR(lower) of lysates of HeLa Tet-Off cells that had been transiently transfected with the specified siRNA (see Supplementary Fig. 3c for data using alternative siRNAs) in the presence of 2 μg/ml of doxycycline (DOX) and, after removing doxycycline 48-h later, with pTRE-FLUC-SOWAHC 3' UTR, which contains a doxycycline-repressible promoter, and the phCMV-MUP reference plasmid. The removal of DOX elicits a transcriptional burst in FLUC-SOWAHC 3' UTR mRNA production. A fraction of cells was harvested after an additional 4-h (time 0) and analyzed. b , Plot of RT-sqPCR data using samples shown in a that were further processed as follows. At time 0, 2 μg/ml of doxycycline were added to the remaining cells. Additional fractions of cells were harvested at the specified times thereafter. RT-sqPCR analyses were essentially as in Fig. 1b. As in Fig. 1d , the levels of exogenous FLUC-SOWAHC 3' UTR mRNAs were comparable to the level of cellular SOWAHC mRNA. Error bars, s.e.m.; # of independently performed experiments = 3; P

    Journal: Nature structural & molecular biology

    Article Title: mRNA-mRNA duplexes that auto-elicit Staufen1-mediated mRNA decay

    doi: 10.1038/nsmb.2664

    Figure Lengend Snippet: mRNA–mRNA duplexes create SBSs and trigger SMD a , Western blot (upper) or RT-sqPCR(lower) of lysates of HeLa Tet-Off cells that had been transiently transfected with the specified siRNA (see Supplementary Fig. 3c for data using alternative siRNAs) in the presence of 2 μg/ml of doxycycline (DOX) and, after removing doxycycline 48-h later, with pTRE-FLUC-SOWAHC 3' UTR, which contains a doxycycline-repressible promoter, and the phCMV-MUP reference plasmid. The removal of DOX elicits a transcriptional burst in FLUC-SOWAHC 3' UTR mRNA production. A fraction of cells was harvested after an additional 4-h (time 0) and analyzed. b , Plot of RT-sqPCR data using samples shown in a that were further processed as follows. At time 0, 2 μg/ml of doxycycline were added to the remaining cells. Additional fractions of cells were harvested at the specified times thereafter. RT-sqPCR analyses were essentially as in Fig. 1b. As in Fig. 1d , the levels of exogenous FLUC-SOWAHC 3' UTR mRNAs were comparable to the level of cellular SOWAHC mRNA. Error bars, s.e.m.; # of independently performed experiments = 3; P

    Article Snippet: Cell culture, transient transfections, formaldehyde crosslinking, and mRNA half-life determinations Human HeLa cells, HeLa Tet-off cells (Clontech), HeLa cells stably expressing IRE-Gl Norm mRNA , or HaCaT cells were grown in DMEM (Gibco-BRL) containing 10% fetal bovine serum (Gibco-BRL) at 37°C and in 5% CO2 .

    Techniques: Western Blot, Transfection, Plasmid Preparation

    An mRNA that duplexes with another mRNA by 3' UTR base-pairing is targeted for SMD provided it is translationally active a , Diagram of IRE-Gl mRNA base-paired with FLUC-AS 3' UTR mRNA, which contains 3' UTR sequences engineered to be complementary to 3' UTR sequences of Gl mRNA. b , Western blot (WB) of lysates of HeLa cells stably expressing IRE-Gl mRNA and transiently expressing FLUC-AS 3' UTR mRNA (+) or, as a negative control, FLUC mRNA (−), the specified siRNA, and the reference MUP mRNA. Cells were cultured in the presence of either hemin or deferoxamine mesylate (Df) for 18-h prior to harvesting. c , Histogram representations of RT-sqPCR analyses of samples analyzed in b . The level of FLUC-AS 3' UTR mRNA was normalized to the level of MUP mRNA ( Supplementary Fig. 4b ), and the normalized level in the presence of Df and Control siRNA is defined as 100. d , as in c , but the level of IRE-Gl mRNA was normalized to the level of cellular SMG7 mRNA ( Supplementary Fig. 4b ). Error bars, s.e.m.; # of independently performed experiments = 3; *, P

    Journal: Nature structural & molecular biology

    Article Title: mRNA-mRNA duplexes that auto-elicit Staufen1-mediated mRNA decay

    doi: 10.1038/nsmb.2664

    Figure Lengend Snippet: An mRNA that duplexes with another mRNA by 3' UTR base-pairing is targeted for SMD provided it is translationally active a , Diagram of IRE-Gl mRNA base-paired with FLUC-AS 3' UTR mRNA, which contains 3' UTR sequences engineered to be complementary to 3' UTR sequences of Gl mRNA. b , Western blot (WB) of lysates of HeLa cells stably expressing IRE-Gl mRNA and transiently expressing FLUC-AS 3' UTR mRNA (+) or, as a negative control, FLUC mRNA (−), the specified siRNA, and the reference MUP mRNA. Cells were cultured in the presence of either hemin or deferoxamine mesylate (Df) for 18-h prior to harvesting. c , Histogram representations of RT-sqPCR analyses of samples analyzed in b . The level of FLUC-AS 3' UTR mRNA was normalized to the level of MUP mRNA ( Supplementary Fig. 4b ), and the normalized level in the presence of Df and Control siRNA is defined as 100. d , as in c , but the level of IRE-Gl mRNA was normalized to the level of cellular SMG7 mRNA ( Supplementary Fig. 4b ). Error bars, s.e.m.; # of independently performed experiments = 3; *, P

    Article Snippet: Cell culture, transient transfections, formaldehyde crosslinking, and mRNA half-life determinations Human HeLa cells, HeLa Tet-off cells (Clontech), HeLa cells stably expressing IRE-Gl Norm mRNA , or HaCaT cells were grown in DMEM (Gibco-BRL) containing 10% fetal bovine serum (Gibco-BRL) at 37°C and in 5% CO2 .

    Techniques: Western Blot, Stable Transfection, Expressing, Negative Control, Cell Culture