brutp  (Millipore)


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  • 99
    Name:
    5 Bromouridine 5 triphosphate sodium salt
    Description:

    Catalog Number:
    b7166
    Price:
    None
    Applications:
    5-Bromouridine 5'-triphosphate (5-BrUTP) is used to measure transcription via labeling of ribonucleic acids (RNA). RNA or cells labeled via 5-BrUTP incorporation may be detected immunologically with antibodies.
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    Structured Review

    Millipore brutp
    5 Bromouridine 5 triphosphate sodium salt

    https://www.bioz.com/result/brutp/product/Millipore
    Average 99 stars, based on 2323 article reviews
    Price from $9.99 to $1999.99
    brutp - by Bioz Stars, 2021-01
    99/100 stars

    Images

    1) Product Images from "Functional Characterization of Negri Bodies (NBs) in Rabies Virus-Infected Cells: Evidence that NBs Are Sites of Viral Transcription and Replication ▿"

    Article Title: Functional Characterization of Negri Bodies (NBs) in Rabies Virus-Infected Cells: Evidence that NBs Are Sites of Viral Transcription and Replication ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00554-09

    Newly synthesized viral RNAs are present inside the viral IBs. (A) BSR cells were infected at an MOI of 3 for 16 h p.i., transfected with BrUTP for 20 min in the presence (+ActD) or absence (-ActD) of ActD. P protein (as a marker of the Negri bodies) was detected by using the rabbit polyclonal anti-P antibody, followed by incubation with Alexa 568-goat anti-rabbit IgG (P). Newly synthesized RNAs were detected by using mouse MAb anti-BrdU antibody (BrUTP) and goat anti-mouse Alexa 488. Colocalization of the viral RNA (vRNA) with the IBs is shown in the merged panel in the presence of the drug (+ActD). In cells untreated with ActD, cellular mRNA is produced in vast excess over vRNA. Thus, for these images, the PMT gain of the confocal laser scanning microscope is lower, so the signal for vRNA is lower than for that shown in the presence of the drug, and specific colocalization is not readily observed. The scale bars correspond to 12 μm. (B) BrUTP signals are specific to RNA produced by rabies virus. BSR cells were infected with the VTF7-3 recombinant virus and cotransfected with plasmids expressing N and P proteins. Cells were then transfected with BrUTP for 20 min in the presence (+ActD) or absence (-ActD) of ActD. P protein and RNA were detected as in panel A. Note that in the absence of the drug, no BrUTP signal is detectable in the IBs formed by N and P proteins, but some RNA signals are detected in the cytoplasmic inclusions, most probably due to vaccinia virus infection (-ActD). In the presence of the drug, no RNA labeling is observed (+ActD).
    Figure Legend Snippet: Newly synthesized viral RNAs are present inside the viral IBs. (A) BSR cells were infected at an MOI of 3 for 16 h p.i., transfected with BrUTP for 20 min in the presence (+ActD) or absence (-ActD) of ActD. P protein (as a marker of the Negri bodies) was detected by using the rabbit polyclonal anti-P antibody, followed by incubation with Alexa 568-goat anti-rabbit IgG (P). Newly synthesized RNAs were detected by using mouse MAb anti-BrdU antibody (BrUTP) and goat anti-mouse Alexa 488. Colocalization of the viral RNA (vRNA) with the IBs is shown in the merged panel in the presence of the drug (+ActD). In cells untreated with ActD, cellular mRNA is produced in vast excess over vRNA. Thus, for these images, the PMT gain of the confocal laser scanning microscope is lower, so the signal for vRNA is lower than for that shown in the presence of the drug, and specific colocalization is not readily observed. The scale bars correspond to 12 μm. (B) BrUTP signals are specific to RNA produced by rabies virus. BSR cells were infected with the VTF7-3 recombinant virus and cotransfected with plasmids expressing N and P proteins. Cells were then transfected with BrUTP for 20 min in the presence (+ActD) or absence (-ActD) of ActD. P protein and RNA were detected as in panel A. Note that in the absence of the drug, no BrUTP signal is detectable in the IBs formed by N and P proteins, but some RNA signals are detected in the cytoplasmic inclusions, most probably due to vaccinia virus infection (-ActD). In the presence of the drug, no RNA labeling is observed (+ActD).

    Techniques Used: Synthesized, Infection, Transfection, Marker, Incubation, Produced, Laser-Scanning Microscopy, Recombinant, Expressing, Labeling

    2) Product Images from "Induction of Stress Granule-Like Structures in Vesicular Stomatitis Virus-Infected Cells"

    Article Title: Induction of Stress Granule-Like Structures in Vesicular Stomatitis Virus-Infected Cells

    Journal: Journal of Virology

    doi: 10.1128/JVI.02305-12

    The formation of VSV-induced SG-like structures is not dependent on a cellular microtubule or microfilament network. (A and B) A microtubule network is dispensable for the formation of VSV-induced SG-like structures. Cells were treated with DMSO (vehicle control) or Noc for 2 h before mock infection (A) or infection with VSV at an MOI of 1 (B). Mock-infected cells treated with Noc were used for IF to detect tubulin (green) and TIA1 (red). VSV-infected cells were harvested at 7 hpi (9 h after Noc treatment), and specific antibodies were used to detect VSV N (green) and TIA1 (red). (C and D) The formation of VSV-induced SG-like structures does not require a microfilament network. Cells were treated with ethanol (vehicle control) or CytoD for 2 h before mock infection (C) or infection with VSV at an MOI of 1 (D). Mock-infected cells treated with CytoD were used for IF to detect actin (green) and TIA1 (red). VSV-infected cells were harvested at 7 hpi (9 h after CytoD treatment), and specific antibodies were used to detect VSV N (green) and TIA1 (red). The secondary antibodies used in these experiments were Alexa Fluor 594-labeled donkey anti-goat IgG for TIA1 (red) and Alexa Fluor 488-labeled donkey anti-mouse IgG for VSV N, tubulin, or actin (green).
    Figure Legend Snippet: The formation of VSV-induced SG-like structures is not dependent on a cellular microtubule or microfilament network. (A and B) A microtubule network is dispensable for the formation of VSV-induced SG-like structures. Cells were treated with DMSO (vehicle control) or Noc for 2 h before mock infection (A) or infection with VSV at an MOI of 1 (B). Mock-infected cells treated with Noc were used for IF to detect tubulin (green) and TIA1 (red). VSV-infected cells were harvested at 7 hpi (9 h after Noc treatment), and specific antibodies were used to detect VSV N (green) and TIA1 (red). (C and D) The formation of VSV-induced SG-like structures does not require a microfilament network. Cells were treated with ethanol (vehicle control) or CytoD for 2 h before mock infection (C) or infection with VSV at an MOI of 1 (D). Mock-infected cells treated with CytoD were used for IF to detect actin (green) and TIA1 (red). VSV-infected cells were harvested at 7 hpi (9 h after CytoD treatment), and specific antibodies were used to detect VSV N (green) and TIA1 (red). The secondary antibodies used in these experiments were Alexa Fluor 594-labeled donkey anti-goat IgG for TIA1 (red) and Alexa Fluor 488-labeled donkey anti-mouse IgG for VSV N, tubulin, or actin (green).

    Techniques Used: Infection, Labeling

    3) Product Images from "Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming"

    Article Title: Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming

    Journal: Molecular Cell

    doi: 10.1016/j.molcel.2014.06.024

    Rapid Genome-wide Transcriptional Reprogramming in the Absence of Protein Synthesis (A) Experimental design for the RNA-seq analysis of newly transcribed mRNAs before and after NT to Xenopus oocytes. BrUTP is used to label newly transcribed RNAs. (B) Venn diagram of genes classified as activated (reprogrammed), continuously transcribed (maintained), or repressed (downregulated) after NT based on log 2 count per million (log 2 CPM donor cell divided by NT). False discovery rate (FDR)
    Figure Legend Snippet: Rapid Genome-wide Transcriptional Reprogramming in the Absence of Protein Synthesis (A) Experimental design for the RNA-seq analysis of newly transcribed mRNAs before and after NT to Xenopus oocytes. BrUTP is used to label newly transcribed RNAs. (B) Venn diagram of genes classified as activated (reprogrammed), continuously transcribed (maintained), or repressed (downregulated) after NT based on log 2 count per million (log 2 CPM donor cell divided by NT). False discovery rate (FDR)

    Techniques Used: Genome Wide, RNA Sequencing Assay

    Related Articles

    Transfection:

    Article Title: Functional Characterization of Negri Bodies (NBs) in Rabies Virus-Infected Cells: Evidence that NBs Are Sites of Viral Transcription and Replication ▿
    Article Snippet: .. Briefly, the infected cells (16 h p.i.) were treated with 15 μg of ActD/ml for 1 h and then transfected with BrUTP (B 7166; Sigma) at a final concentration of 10 mM by using Lipofectamine 2000 (Invitrogen) and maintained in the presence of ActD for 20 min. .. P protein was detected by using the rabbit polyclonal anti-P antibody, followed by incubation with Alexa 568-conjugated goat anti-rabbit IgG.

    In Vitro:

    Article Title: Long noncoding RNA DRAIC inhibits prostate cancer progression by interacting with IKK to inhibit NF-κB activation
    Article Snippet: .. The sense and antisense DRAIC RNA was labeled by random incorporation of 5-Bromo-UTP during the in vitro transcription. .. 5 μg of each sense and antisense in vitro transcribed DRAIC RNA was used per pull down experiment.

    Immunoprecipitation:

    Article Title: Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming
    Article Snippet: .. RNA Immunoprecipitation BrUTP (Sigma: B7166, 4.6 nl of 100 mM stock) was injected to the cytoplasm of Xenopus oocytes 2 hr after transplantation with MEFs. .. Oocytes were collected 48 hr after NT, and RNA was extracted using a QIAGEN RNeasy kit (eight oocytes per column).

    Synthesized:

    Article Title: Identification of Severe Acute Respiratory Syndrome Coronavirus Replicase Products and Characterization of Papain-Like Protease Activity
    Article Snippet: .. Newly synthesized viral RNA was labeled with 5-bromouridine 5′-triphosphate (BrUTP; Sigma, St Louis, Mo.) as previously described for mouse hepatitis virus infection ( ). .. Briefly, Vero E6 cells were added to chamber slides, and when the cells were ≈60% confluent, they were mock infected or infected with 1,000 TCID50 of SARS coronavirus and incubated in complete medium.

    Infection:

    Article Title: Functional Characterization of Negri Bodies (NBs) in Rabies Virus-Infected Cells: Evidence that NBs Are Sites of Viral Transcription and Replication ▿
    Article Snippet: .. Briefly, the infected cells (16 h p.i.) were treated with 15 μg of ActD/ml for 1 h and then transfected with BrUTP (B 7166; Sigma) at a final concentration of 10 mM by using Lipofectamine 2000 (Invitrogen) and maintained in the presence of ActD for 20 min. .. P protein was detected by using the rabbit polyclonal anti-P antibody, followed by incubation with Alexa 568-conjugated goat anti-rabbit IgG.

    Article Title: Identification of Severe Acute Respiratory Syndrome Coronavirus Replicase Products and Characterization of Papain-Like Protease Activity
    Article Snippet: .. Newly synthesized viral RNA was labeled with 5-bromouridine 5′-triphosphate (BrUTP; Sigma, St Louis, Mo.) as previously described for mouse hepatitis virus infection ( ). .. Briefly, Vero E6 cells were added to chamber slides, and when the cells were ≈60% confluent, they were mock infected or infected with 1,000 TCID50 of SARS coronavirus and incubated in complete medium.

    Concentration Assay:

    Article Title: Functional Characterization of Negri Bodies (NBs) in Rabies Virus-Infected Cells: Evidence that NBs Are Sites of Viral Transcription and Replication ▿
    Article Snippet: .. Briefly, the infected cells (16 h p.i.) were treated with 15 μg of ActD/ml for 1 h and then transfected with BrUTP (B 7166; Sigma) at a final concentration of 10 mM by using Lipofectamine 2000 (Invitrogen) and maintained in the presence of ActD for 20 min. .. P protein was detected by using the rabbit polyclonal anti-P antibody, followed by incubation with Alexa 568-conjugated goat anti-rabbit IgG.

    Article Title: Nonphosphorylated Human La Antigen Interacts with Nucleolin at Nucleolar Sites Involved in rRNA Biogenesis
    Article Snippet: .. Cells were washed, permeabilized, and incubated for 10 min at 37°C in transcription buffer containing 0.5 mM 5-bromouridine 5′-triphosphate (BrUTP; Sigma) containing the appropriate concentration of α-amanitin and processed as described previously with primary monoclonal Ab against BrUTP (1:200; Caltag) and AbNP (1:50) in phosphate-buffered saline for 1 h, washed with phosphate-buffered saline, and incubated with secondary Ab conjugated with fluorescein or Texas red (Vector Laboratories) ( , ). .. Fluorescence resonance energy transfer (FRET) experiments were performed in transfected HeLa cells on a Zeiss 510 confocal microscope with a ×100 and 1.3 numerical aperture planapochromat oil objective and ×3 zoom.

    Incubation:

    Article Title: Chromosome intermingling—the physical basis of chromosome organization in differentiated cells
    Article Snippet: .. This was followed by incubation with 10 mM Br-UTP (Sigma, B7166) in hypotonic buffer at room temperature. .. After 5 min, the hypotonic media was replaced with 10% FBS containing DMEM medium and incubated at 37°C and 5% CO2 for 20 min before fixing it with freshly thawed 4% PFA (in PBS) for 10 min.

    Article Title: Nonphosphorylated Human La Antigen Interacts with Nucleolin at Nucleolar Sites Involved in rRNA Biogenesis
    Article Snippet: .. Cells were washed, permeabilized, and incubated for 10 min at 37°C in transcription buffer containing 0.5 mM 5-bromouridine 5′-triphosphate (BrUTP; Sigma) containing the appropriate concentration of α-amanitin and processed as described previously with primary monoclonal Ab against BrUTP (1:200; Caltag) and AbNP (1:50) in phosphate-buffered saline for 1 h, washed with phosphate-buffered saline, and incubated with secondary Ab conjugated with fluorescein or Texas red (Vector Laboratories) ( , ). .. Fluorescence resonance energy transfer (FRET) experiments were performed in transfected HeLa cells on a Zeiss 510 confocal microscope with a ×100 and 1.3 numerical aperture planapochromat oil objective and ×3 zoom.

    Labeling:

    Article Title: Long noncoding RNA DRAIC inhibits prostate cancer progression by interacting with IKK to inhibit NF-κB activation
    Article Snippet: .. The sense and antisense DRAIC RNA was labeled by random incorporation of 5-Bromo-UTP during the in vitro transcription. .. 5 μg of each sense and antisense in vitro transcribed DRAIC RNA was used per pull down experiment.

    Article Title: Identification of Severe Acute Respiratory Syndrome Coronavirus Replicase Products and Characterization of Papain-Like Protease Activity
    Article Snippet: .. Newly synthesized viral RNA was labeled with 5-bromouridine 5′-triphosphate (BrUTP; Sigma, St Louis, Mo.) as previously described for mouse hepatitis virus infection ( ). .. Briefly, Vero E6 cells were added to chamber slides, and when the cells were ≈60% confluent, they were mock infected or infected with 1,000 TCID50 of SARS coronavirus and incubated in complete medium.

    Transplantation Assay:

    Article Title: Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming
    Article Snippet: .. RNA Immunoprecipitation BrUTP (Sigma: B7166, 4.6 nl of 100 mM stock) was injected to the cytoplasm of Xenopus oocytes 2 hr after transplantation with MEFs. .. Oocytes were collected 48 hr after NT, and RNA was extracted using a QIAGEN RNeasy kit (eight oocytes per column).

    Injection:

    Article Title: Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming
    Article Snippet: .. RNA Immunoprecipitation BrUTP (Sigma: B7166, 4.6 nl of 100 mM stock) was injected to the cytoplasm of Xenopus oocytes 2 hr after transplantation with MEFs. .. Oocytes were collected 48 hr after NT, and RNA was extracted using a QIAGEN RNeasy kit (eight oocytes per column).

    Article Title: Gene Resistance to Transcriptional Reprogramming following Nuclear Transfer Is Directly Mediated by Multiple Chromatin-Repressive Pathways
    Article Snippet: .. Using a Drummond injector (Drummond Nanoject, Drummond Scientific Company, USA), the required amount of mRNAs or gRNAs (9nl at 1mg/ml), or BrUTP (Sigma: B7166, 4.6nl at 100mM), is injected in the cytoplasm from the vegetal pole of the recipient oocyte. .. Injected oocytes are cultured in 1X MBS, P/S, 0.2%BSA at 18°C.

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  • 95
    Millipore 5 bromo utp
    5 Bromo Utp, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5 bromo utp/product/Millipore
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    5 bromo utp - by Bioz Stars, 2021-01
    95/100 stars
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