5 bromouridine 5 triphosphate brutp  (Millipore)

 
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    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 5 bromouridine 5 triphosphate brutp
    5 Bromouridine 5 triphosphate sodium salt

    https://www.bioz.com/result/5 bromouridine 5 triphosphate brutp/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    5 bromouridine 5 triphosphate brutp - by Bioz Stars, 2021-03
    93/100 stars

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    Related Articles

    Incubation:

    Article Title: Hrp59, an hnRNP M protein in Chironomus and Drosophila, binds to exonic splicing enhancers and is required for expression of a subset of mRNAs
    Article Snippet: Glands were subsequently washed for 3 min in 0.5% Triton X-100/TBS, 5 min in TBS, and fixed and coimmunostained with an mAb against bromo-uridine (Roche) and the polyclonal anti-Hrp59, as described above for immunofluorescent labeling of salivary gland cells. .. RNase treatment of C. tentans salivary glands Salivary glands were dissected and preextracted as for BrUTP incorporation, washed in TBS and incubated for 30 min with 0.1 mg/ml DNase-free RNase A (Sigma-Aldrich) in TBS, or with TBS alone. .. The glands were then fixed and immunostained using antibodies against either Hrp65 or Hrp59 as described for immunofluorescent labeling of salivary gland cells.

    Article Title: Spatio-temporal Dynamics of Replication and Transcription Sites in the Mammalian Cell Nucleus
    Article Snippet: Exponentially growing HeLa cells on glass cover slips were pulsed with CldU (20 μM) for 7 min to label replication sites and then washed with ice-cold TBS buffer (10 mM Tris-HCl, pH 7.4, 150 mM NaCl and 5 mM MgCl2 ), followed by glycerol buffer (20 mM Tris-HCl, pH 7.4, 25% glycerol, 5 mM MgCl2, 0.5 mM EGTA, 0.5 mM PMSF) for 10 min on ice. .. Washed cells were permeabilized with 0.025% Triton X-100 in glycerol buffer (with 25 U/ml of RNasin; Promega Corp.) on ice for 3 min and immediately incubated at room temperature for 30 min with nucleic acid synthesis buffer (50 mM Tris-HCl, pH 7.4, 10 mM MgCl2, 150 mM NaCl, 25% glycerol, 0.5 mM PMSF, 25 U/ml of RNasin (Promega), 1.8 mM ATP) supplemented with 0.5 mM CTP, GTP, and BrUTP (Sigma Chemical Co) to label transcription sites. ..

    Article Title: Implication of Localization of Human DNA Repair Enzyme O6-Methylguanine-DNA Methyltransferase at Active Transcription Sites in Transcription-Repair Coupling of the Mutagenic O6-Methylguanine Lesion
    Article Snippet: When one-third confluency was reached, cells were incubated with the required media for various times: (i) normal growth condition (G1 ) with 10% fetal bovine serum (FBS) in minimal essential medium (MEM), (ii) G0 with 1% FBS followed by 0% FBS in MEM, and (iii) S with 2.5 mM thymidine in MEM with 10% FBS. .. For labelling of pre-mRNA by BrUTP, streptolysin O-permeabilized agarose-encapsulated cells were incubated in a mixture containing 2 mM ATP; 0.1 mM CTP, GTP, and UTP (or BrUTP [Sigma]); and 2 mM MgCl2 as described previously ( , ). .. After fixation with 4% paraformaldehyde, cells were stained with MGMT.PAb and MAb.BrdU before they were spun onto slides for microscopy.

    Article Title: Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation
    Article Snippet: Next, one unit of reverse transcriptase (Expand RT, Roche Molecular Biochemicals) and one unit of RNase inhibitor (Promega) were added, and extension was performed at 42°C for 90 minutes. .. Microscopy Labeling of nascent RNA using BrUTP was performed essentially according to , only the Saponin (Sigma) incubation was performed at 4°C. .. After the transcription reaction, cells were washed and fixed in 2% paraformaldehyde and BrUTP labeled transcripts were detected with a monoclonal anti-BrdUTP antibody (Roche).

    Transfection:

    Article Title: Bluetongue virus RNA binding protein NS2 is a modulator of viral replication and assembly
    Article Snippet: Detection of newly synthesized RNA in vivo by IF BSR cells were infected with BTV at MOI of 1 for 14 h, followed by treatment with actinomycin D (10 μg/ml). .. After 1 h cells were transfected with 10 mM BrUTP (Sigma) using 5% FuGene transfection reagent (Roche). .. Cells were fixed 17 h pi and processed for IF analysis with mouse anti-bromodeoxyuridine BrdU monoclonal antisera (Sigma) followed by Alexa Fluor 594 goat anti-mouse IgG as secondary antibody.

    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.

    Microscopy:

    Article Title: Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation
    Article Snippet: Next, one unit of reverse transcriptase (Expand RT, Roche Molecular Biochemicals) and one unit of RNase inhibitor (Promega) were added, and extension was performed at 42°C for 90 minutes. .. Microscopy Labeling of nascent RNA using BrUTP was performed essentially according to , only the Saponin (Sigma) incubation was performed at 4°C. .. After the transcription reaction, cells were washed and fixed in 2% paraformaldehyde and BrUTP labeled transcripts were detected with a monoclonal anti-BrdUTP antibody (Roche).

    Labeling:

    Article Title: Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation
    Article Snippet: Next, one unit of reverse transcriptase (Expand RT, Roche Molecular Biochemicals) and one unit of RNase inhibitor (Promega) were added, and extension was performed at 42°C for 90 minutes. .. Microscopy Labeling of nascent RNA using BrUTP was performed essentially according to , only the Saponin (Sigma) incubation was performed at 4°C. .. After the transcription reaction, cells were washed and fixed in 2% paraformaldehyde and BrUTP labeled transcripts were detected with a monoclonal anti-BrdUTP antibody (Roche).

    Article Title: Localization of Mouse Hepatitis Virus Nonstructural Proteins and RNA Synthesis Indicates a Role for Late Endosomes in Viral Replication
    Article Snippet: .. The metabolic labeling of viral RNA synthesis with BrUTP (Sigma) has been described recently ( ). .. Briefly, MHV-infected L cells were given 10 μg of dactinomycin (Sigma)/ml 30 min prior to labeling.

    Immunoprecipitation:

    Article Title: Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming
    Article Snippet: Real-time PCR was performed as SYBR Green assays on an ABI 7300 Real Time PCR Cycler using a standard ABI cycling condition. .. 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: Real-time PCR was performed as SYBR Green assays on an ABI 7300 Real Time PCR Cycler using a standard ABI cycling condition. .. 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).

    Transplantation Assay:

    Article Title: Hierarchical Molecular Events Driven by Oocyte-Specific Factors Lead to Rapid and Extensive Reprogramming
    Article Snippet: Real-time PCR was performed as SYBR Green assays on an ABI 7300 Real Time PCR Cycler using a standard ABI cycling condition. .. 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).

    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.

    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.

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  • 93
    Millipore brutp
    Colocalization of incorporated BrUMP and MGMT speckles. (a) <t>BrUTP</t> with agarose-encapsulated cells. Pulse-labelling of newly synthesized RNA (or active-transcription sites) by incubating streptolysin <t>O-permeabilized</t> agarose-embedded cells with transcription labelling mixture containing BrUTP. Magnification, ca. ×40. A and B, C and D, and E and F are identical cells stained by MGMT.PAb (in red) and MAb-BrdU (in green; for incorporated BrUMP) simultaneously; note that the cells are spherical in shape due to the embedded agarose. (b) BrUTP with microinjection. Transcription labelling mixture in phosphate-buffered saline was injected into the nuclei of HeLa.CCL2B cells. Injected cells were incubated at 37°C for the required time in minutes before fixation with 4% paraformaldehyde for staining similar to that described for panel a. Magnification, ca. ×90. A and B, C and D, E and F, G and H, I and J, and K and L are identical cells visualized by single-wavelength excitation for MGMT in red and incorporated BrUMP in green, respectively. Pictures K and L are control cells injected with transcription labelling mixture containing UTP instead of BrUTP. (c) Dual-antigen stainings. MGMT and incorporated BrUMP speckles (after 10 min of labelling) were visualized by single (red or green)- and double (“Double exc.” in picture D)-wavelength excitations for half the exposure time of panel c.
    Brutp, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/brutp/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    brutp - by Bioz Stars, 2021-03
    93/100 stars
      Buy from Supplier

    Image Search Results


    Colocalization of incorporated BrUMP and MGMT speckles. (a) BrUTP with agarose-encapsulated cells. Pulse-labelling of newly synthesized RNA (or active-transcription sites) by incubating streptolysin O-permeabilized agarose-embedded cells with transcription labelling mixture containing BrUTP. Magnification, ca. ×40. A and B, C and D, and E and F are identical cells stained by MGMT.PAb (in red) and MAb-BrdU (in green; for incorporated BrUMP) simultaneously; note that the cells are spherical in shape due to the embedded agarose. (b) BrUTP with microinjection. Transcription labelling mixture in phosphate-buffered saline was injected into the nuclei of HeLa.CCL2B cells. Injected cells were incubated at 37°C for the required time in minutes before fixation with 4% paraformaldehyde for staining similar to that described for panel a. Magnification, ca. ×90. A and B, C and D, E and F, G and H, I and J, and K and L are identical cells visualized by single-wavelength excitation for MGMT in red and incorporated BrUMP in green, respectively. Pictures K and L are control cells injected with transcription labelling mixture containing UTP instead of BrUTP. (c) Dual-antigen stainings. MGMT and incorporated BrUMP speckles (after 10 min of labelling) were visualized by single (red or green)- and double (“Double exc.” in picture D)-wavelength excitations for half the exposure time of panel c.

    Journal: Molecular and Cellular Biology

    Article Title: Implication of Localization of Human DNA Repair Enzyme O6-Methylguanine-DNA Methyltransferase at Active Transcription Sites in Transcription-Repair Coupling of the Mutagenic O6-Methylguanine Lesion

    doi:

    Figure Lengend Snippet: Colocalization of incorporated BrUMP and MGMT speckles. (a) BrUTP with agarose-encapsulated cells. Pulse-labelling of newly synthesized RNA (or active-transcription sites) by incubating streptolysin O-permeabilized agarose-embedded cells with transcription labelling mixture containing BrUTP. Magnification, ca. ×40. A and B, C and D, and E and F are identical cells stained by MGMT.PAb (in red) and MAb-BrdU (in green; for incorporated BrUMP) simultaneously; note that the cells are spherical in shape due to the embedded agarose. (b) BrUTP with microinjection. Transcription labelling mixture in phosphate-buffered saline was injected into the nuclei of HeLa.CCL2B cells. Injected cells were incubated at 37°C for the required time in minutes before fixation with 4% paraformaldehyde for staining similar to that described for panel a. Magnification, ca. ×90. A and B, C and D, E and F, G and H, I and J, and K and L are identical cells visualized by single-wavelength excitation for MGMT in red and incorporated BrUMP in green, respectively. Pictures K and L are control cells injected with transcription labelling mixture containing UTP instead of BrUTP. (c) Dual-antigen stainings. MGMT and incorporated BrUMP speckles (after 10 min of labelling) were visualized by single (red or green)- and double (“Double exc.” in picture D)-wavelength excitations for half the exposure time of panel c.

    Article Snippet: For labelling of pre-mRNA by BrUTP, streptolysin O-permeabilized agarose-encapsulated cells were incubated in a mixture containing 2 mM ATP; 0.1 mM CTP, GTP, and UTP (or BrUTP [Sigma]); and 2 mM MgCl2 as described previously ( , ).

    Techniques: Synthesized, Staining, Injection, Incubation

    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).

    Journal: Journal of Virology

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

    doi: 10.1128/JVI.00554-09

    Figure Lengend 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).

    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.

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

    Transcription analysis of cells where VSG synthesis is blocked. A) Northern blot analysis of T. brucei 221VG1.1 cells where VSG221 RNAi had been induced for the time indicated above in hours (h). The parental (P) T. brucei 90-13 cell line does not contain the VSG221 RNAi construct. The T. brucei 221VG1.1 cell line was either not incubated with tetracycline (− Tet) or had VSG221 RNAi induced with tetracycline for the times indicated above. Blots were hybridised with probes for Actin, NUP1 , eGFP (present in the active VSG221 ES), TDP1 , tubulin (Tub), ESAG5 , 18S rRNA, ISWI , ESAG6/7, VSG221 and PFR2 . Ethidium stains of the gels are indicated on the right to indicate total RNA loaded. B) Transcription analysis of T. brucei 221VG1.1 where VSG synthesis was blocked by the induction of VSG221 RNAi with tetracycline (Tet) for 0 or 24 hours (h). Cells were incubated with BrUTP to label nascent transcripts and subsequently incubated with an anti-BrdUTP antibody, and a secondary antibody coupled to Alexa 488. DNA was stained with DAPI. A normal precytokinesis cell (0 h) is compared with a precytokinesis cell arising after the induction of VSG RNAi for 24 hours (24 h). The experiment was performed in the absence of α-amanitin (− α-ama) to visualise total transcription, or in the presence of 200 µg ml −1 α-amanitin to inhibit transcription by RNA polymerases II and III and visualise transcription by RNA polymerase I (+ α-ama). The scale bar indicates 4 µm. Quantitation of transcription as fluorescence in the FITC channel is in arbitrary units using 50 cells per time point, with standard deviation indicated with error bars.

    Journal: PLoS ONE

    Article Title: Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation

    doi: 10.1371/journal.pone.0007532

    Figure Lengend Snippet: Transcription analysis of cells where VSG synthesis is blocked. A) Northern blot analysis of T. brucei 221VG1.1 cells where VSG221 RNAi had been induced for the time indicated above in hours (h). The parental (P) T. brucei 90-13 cell line does not contain the VSG221 RNAi construct. The T. brucei 221VG1.1 cell line was either not incubated with tetracycline (− Tet) or had VSG221 RNAi induced with tetracycline for the times indicated above. Blots were hybridised with probes for Actin, NUP1 , eGFP (present in the active VSG221 ES), TDP1 , tubulin (Tub), ESAG5 , 18S rRNA, ISWI , ESAG6/7, VSG221 and PFR2 . Ethidium stains of the gels are indicated on the right to indicate total RNA loaded. B) Transcription analysis of T. brucei 221VG1.1 where VSG synthesis was blocked by the induction of VSG221 RNAi with tetracycline (Tet) for 0 or 24 hours (h). Cells were incubated with BrUTP to label nascent transcripts and subsequently incubated with an anti-BrdUTP antibody, and a secondary antibody coupled to Alexa 488. DNA was stained with DAPI. A normal precytokinesis cell (0 h) is compared with a precytokinesis cell arising after the induction of VSG RNAi for 24 hours (24 h). The experiment was performed in the absence of α-amanitin (− α-ama) to visualise total transcription, or in the presence of 200 µg ml −1 α-amanitin to inhibit transcription by RNA polymerases II and III and visualise transcription by RNA polymerase I (+ α-ama). The scale bar indicates 4 µm. Quantitation of transcription as fluorescence in the FITC channel is in arbitrary units using 50 cells per time point, with standard deviation indicated with error bars.

    Article Snippet: Microscopy Labeling of nascent RNA using BrUTP was performed essentially according to , only the Saponin (Sigma) incubation was performed at 4°C.

    Techniques: Northern Blot, Construct, Incubation, Staining, Quantitation Assay, Fluorescence, Standard Deviation

    Distribution of Hrp59 and Hrp65 in the polytene chromosomes. (A) Squash preparations of C. tentans polytene chromosomes were double labeled with antibodies against Hrp59 (green) and Hrp65 (red). The BR1 and BR2 puffs in chromosome IV are indicated by arrowheads. The large arrow points to a band with intense labeling for both Hrp59 and Hrp65. The short arrows point to bands labeled by only one of the two antibodies. Note that the distributions of Hrp59 and Hrp65 are similar. (B) Polytene chromosome double labeled with antibodies against Hrp59 (green) and Hrp45 (red). The arrowheads point to the BR1 and BR2 puffs. Hrp45 and Hrp59 show different labeling patterns. (C) Transcriptionally active loci were visualized by BrUTP incorporation (red). The glands were coimmunostained with anti-Hrp59 antibody (green). C, cytoplasm; N, nucleolus; NP, nucleoplasm; PC, polytene chromosome. (D) The distributions of Hrp59 (green) and Hrp65 (red) in the polytene chromosomes were analyzed after heat shock. Note the redistribution of both proteins to the heat-shock locus at 5C (compare chromosome IV in A and D). (E) Tissue culture cells of C. tentans , either nontreated (22°C) or heat shocked (37°C), were double stained with anti-Hrp59 (green) and anti-Hrp65 (red) antibodies. All images correspond to optical sections with a thickness of ∼1 μm. Bars: (A, B, and D) ∼10 μm; (C) 20 μm; (E) 5 μm.

    Journal: The Journal of Cell Biology

    Article Title: Hrp59, an hnRNP M protein in Chironomus and Drosophila, binds to exonic splicing enhancers and is required for expression of a subset of mRNAs

    doi: 10.1083/jcb.200407173

    Figure Lengend Snippet: Distribution of Hrp59 and Hrp65 in the polytene chromosomes. (A) Squash preparations of C. tentans polytene chromosomes were double labeled with antibodies against Hrp59 (green) and Hrp65 (red). The BR1 and BR2 puffs in chromosome IV are indicated by arrowheads. The large arrow points to a band with intense labeling for both Hrp59 and Hrp65. The short arrows point to bands labeled by only one of the two antibodies. Note that the distributions of Hrp59 and Hrp65 are similar. (B) Polytene chromosome double labeled with antibodies against Hrp59 (green) and Hrp45 (red). The arrowheads point to the BR1 and BR2 puffs. Hrp45 and Hrp59 show different labeling patterns. (C) Transcriptionally active loci were visualized by BrUTP incorporation (red). The glands were coimmunostained with anti-Hrp59 antibody (green). C, cytoplasm; N, nucleolus; NP, nucleoplasm; PC, polytene chromosome. (D) The distributions of Hrp59 (green) and Hrp65 (red) in the polytene chromosomes were analyzed after heat shock. Note the redistribution of both proteins to the heat-shock locus at 5C (compare chromosome IV in A and D). (E) Tissue culture cells of C. tentans , either nontreated (22°C) or heat shocked (37°C), were double stained with anti-Hrp59 (green) and anti-Hrp65 (red) antibodies. All images correspond to optical sections with a thickness of ∼1 μm. Bars: (A, B, and D) ∼10 μm; (C) 20 μm; (E) 5 μm.

    Article Snippet: RNase treatment of C. tentans salivary glands Salivary glands were dissected and preextracted as for BrUTP incorporation, washed in TBS and incubated for 30 min with 0.1 mg/ml DNase-free RNase A (Sigma-Aldrich) in TBS, or with TBS alone.

    Techniques: Labeling, Staining