brdu  (Thermo Fisher)


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    Name:
    BrdU 5 Bromo 2 Deoxyuridine
    Description:
    Cells can naturally incorporate the thymidine analog BrdU into their cells during cell division making this nucleoside analog an excellent marker of by cell cycle and proliferation Analysis of incorporated BrdU can be with an anti BrdU antibody
    Catalog Number:
    B23151
    Price:
    None
    Category:
    Oligos Primers Probes Nucleotides
    Applications:
    Apoptosis|Cell Analysis|Cell Cycle|Cell Proliferation|DNA Fragmentation|DNA Fragmentation Assays|Flow Cytometry Apoptosis Assays|Flow Cytometry Cell Proliferation|Flow Cytometry of Cellular Processes|Flow Cytometry|Cell Viability, Proliferation & Function
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    Structured Review

    Thermo Fisher brdu
    Effect of Par2b deficiency on increased <t>BrdU</t> incorporation associated with loss of hai1a function in the basal layer versus periderm. (A and B) Control and hai1a morphants periderm-nuclear-EGFP embryos with and without St14a or Par2b deficiency were incubated with BrdU for 3 h beginning at 28 <t>hpf,</t> collected for analysis at 46 hpf, and immunostained for p63 and BrdU. Bar, 50 µm. (B) Colocation of BrdU staining with either nuclear-GFP or nuclear p63 staining was used to identify periderm or basal layer cells that had incorporated BrdU, respectively. (C and D) The percentage of BrdU-positive nuclei in periderm (C) and basal layer (D) is shown (mean ± SEM of three experiments; for each experiment, 10 embryos were examined per condition, and 150–200 cells were analyzed in each embryo). Results were analyzed by one-way ANOVA and Bonferroni posttest. In C, the hai1a/st14a MO group and the par2b −/− hai1a/st14a MO groups were not different from control; the hai1a MO group, the par2b −/− group, and the par2b −/− hai1a MO group were all different from control, and the par2b −/− and hai1a MO groups were different from the par2b −/− hai1a MO group (*, P
    Cells can naturally incorporate the thymidine analog BrdU into their cells during cell division making this nucleoside analog an excellent marker of by cell cycle and proliferation Analysis of incorporated BrdU can be with an anti BrdU antibody
    https://www.bioz.com/result/brdu/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    brdu - by Bioz Stars, 2021-05
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    Images

    1) Product Images from "Protease signaling regulates apical cell extrusion, cell contacts, and proliferation in epithelia"

    Article Title: Protease signaling regulates apical cell extrusion, cell contacts, and proliferation in epithelia

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201709118

    Effect of Par2b deficiency on increased BrdU incorporation associated with loss of hai1a function in the basal layer versus periderm. (A and B) Control and hai1a morphants periderm-nuclear-EGFP embryos with and without St14a or Par2b deficiency were incubated with BrdU for 3 h beginning at 28 hpf, collected for analysis at 46 hpf, and immunostained for p63 and BrdU. Bar, 50 µm. (B) Colocation of BrdU staining with either nuclear-GFP or nuclear p63 staining was used to identify periderm or basal layer cells that had incorporated BrdU, respectively. (C and D) The percentage of BrdU-positive nuclei in periderm (C) and basal layer (D) is shown (mean ± SEM of three experiments; for each experiment, 10 embryos were examined per condition, and 150–200 cells were analyzed in each embryo). Results were analyzed by one-way ANOVA and Bonferroni posttest. In C, the hai1a/st14a MO group and the par2b −/− hai1a/st14a MO groups were not different from control; the hai1a MO group, the par2b −/− group, and the par2b −/− hai1a MO group were all different from control, and the par2b −/− and hai1a MO groups were different from the par2b −/− hai1a MO group (*, P
    Figure Legend Snippet: Effect of Par2b deficiency on increased BrdU incorporation associated with loss of hai1a function in the basal layer versus periderm. (A and B) Control and hai1a morphants periderm-nuclear-EGFP embryos with and without St14a or Par2b deficiency were incubated with BrdU for 3 h beginning at 28 hpf, collected for analysis at 46 hpf, and immunostained for p63 and BrdU. Bar, 50 µm. (B) Colocation of BrdU staining with either nuclear-GFP or nuclear p63 staining was used to identify periderm or basal layer cells that had incorporated BrdU, respectively. (C and D) The percentage of BrdU-positive nuclei in periderm (C) and basal layer (D) is shown (mean ± SEM of three experiments; for each experiment, 10 embryos were examined per condition, and 150–200 cells were analyzed in each embryo). Results were analyzed by one-way ANOVA and Bonferroni posttest. In C, the hai1a/st14a MO group and the par2b −/− hai1a/st14a MO groups were not different from control; the hai1a MO group, the par2b −/− group, and the par2b −/− hai1a MO group were all different from control, and the par2b −/− and hai1a MO groups were different from the par2b −/− hai1a MO group (*, P

    Techniques Used: BrdU Incorporation Assay, Incubation, BrdU Staining, Staining

    2) Product Images from "aP2-Cre-mediated inactivation of acetyl-CoA carboxylase 1 causes growth retardation and reduced lipid accumulation in adipose tissues"

    Article Title: aP2-Cre-mediated inactivation of acetyl-CoA carboxylase 1 causes growth retardation and reduced lipid accumulation in adipose tissues

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0909055106

    Reduced chondrocyte proliferation and bone formation in FACC1KO mice. ( A ) Paraffin-embedded nondecalcified sections from tibias of 2-day-old WT mice ( n = 6) and their FACC1KO littermates ( n = 3) were immunostained for BrdU. BrdU positive cells have brown
    Figure Legend Snippet: Reduced chondrocyte proliferation and bone formation in FACC1KO mice. ( A ) Paraffin-embedded nondecalcified sections from tibias of 2-day-old WT mice ( n = 6) and their FACC1KO littermates ( n = 3) were immunostained for BrdU. BrdU positive cells have brown

    Techniques Used: Mouse Assay

    3) Product Images from "Apoptotic effect of novel Schiff Based CdCl2(C14H21N3O2) complex is mediated via activation of the mitochondrial pathway in colon cancer cells"

    Article Title: Apoptotic effect of novel Schiff Based CdCl2(C14H21N3O2) complex is mediated via activation of the mitochondrial pathway in colon cancer cells

    Journal: Scientific Reports

    doi: 10.1038/srep09097

    Activity of CdCl 2 (C 14 H 21 N 3 O 2 ) complex on cell cycle arrest in the S/M phase. (A) After incubation with DMSO (negative control) or Schiff based compound (3 μg/mL) for 24 h, HT-29 cells were collected, stained with BrdU (representing the S phase) and Phospho-Histone H3 (representing the M phase), and subjected to cell cycle analysis using a Cellomics ArrayScan HCS reader. (B) Representative bar charts showing that treatment with CdCl 2 (C 14 H 21 N 3 O 2 ) complex caused no significant changes in the BrdU and Phospho-Histone H3 fluorescence intensities, suggesting that the cells do not arrest at the S/M phases. The data represented the means ± SD of the fluorescence intensity readings from three independent experiments. * P
    Figure Legend Snippet: Activity of CdCl 2 (C 14 H 21 N 3 O 2 ) complex on cell cycle arrest in the S/M phase. (A) After incubation with DMSO (negative control) or Schiff based compound (3 μg/mL) for 24 h, HT-29 cells were collected, stained with BrdU (representing the S phase) and Phospho-Histone H3 (representing the M phase), and subjected to cell cycle analysis using a Cellomics ArrayScan HCS reader. (B) Representative bar charts showing that treatment with CdCl 2 (C 14 H 21 N 3 O 2 ) complex caused no significant changes in the BrdU and Phospho-Histone H3 fluorescence intensities, suggesting that the cells do not arrest at the S/M phases. The data represented the means ± SD of the fluorescence intensity readings from three independent experiments. * P

    Techniques Used: Activity Assay, Incubation, Negative Control, Staining, Cell Cycle Assay, Fluorescence

    4) Product Images from "MYC Gene Delivery to Adult Mouse Utricles Stimulates Proliferation of Postmitotic Supporting Cells In Vitro"

    Article Title: MYC Gene Delivery to Adult Mouse Utricles Stimulates Proliferation of Postmitotic Supporting Cells In Vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0048704

    Supporting cells that reenter the cell cycle after Ad.MT58A infection can progress to mitosis. ( A ) Confocal images show an Ad.MT58A-infected utricle (1×10 9 TU/mL) that was fixed at 7 days post-virus (DPV) and co-labeled with antibodies to BrdU (red) and Ki-67 (green). Phalloidin labeling (grayscale) is shown to aid in visualizing the borders of the sensory epithelium. White dashed lines demarcate the borders of the sensory epithelium. Scale bar, 100 µm. Insets show high-resolution views of nuclei in the sensory epithelium. Arrows indicate nuclei that labeled with antibodies to BrdU but not Ki67. Scale bar for insets, 5 µm. ( B ) Graph shows the mean number of BrdU-labeled nuclei (green data points) and Ki-67-labeled nuclei (red data points) per sensory epithelium at 5, 7, and 10 DPV. ( C ) Graph shows quantification of the percentage of the BrdU-positive population that did not label with Ki67 antibodies (green data points) and the percentage of the Ki-67-positive population that did not label with BrdU antibodies (red data points). ( D ) Confocal image of an adult mouse utricle infected with Ad.MT58A (1×10 9 TU/mL) that was fixed at 7 DPV and co-labeled with antibodies to PH3-Ser10 (white) and Ki-67 (red). ( E ) Confocal images of a utricle from an embryonic day 17.5 (E17.5) mouse that was fixed in vivo and co-labeled with antibodies to PH3-Ser10 (white) and Ki-67 (red). Phalloidin labeling (grayscale) is shown to aid in visualizing the borders of the sensory epithelium. White dashed lines demarcate the borders of the sensory epithelium, and arrows in D and E indicate PH3-Ser10/Ki-67 co-labeled nuclei. Scale bar for D–E, 100 µm. ( F ) Graph shows quantification of the percentage of the Ki-67-positive populations that labeled with antibodies to PH3-Ser10. The difference in the percentage of PH3-Ser10-positive/Ki-67-positive nuclei did not reach statistical significance (p > 0.05; Student's t-test). The numbers above the gray bars indicate the mean number of PH3-Ser10-labeled nuclei per sensory epithelium. There were significantly more PH3-Ser10-positive nuclei in E17.5 utricles (p
    Figure Legend Snippet: Supporting cells that reenter the cell cycle after Ad.MT58A infection can progress to mitosis. ( A ) Confocal images show an Ad.MT58A-infected utricle (1×10 9 TU/mL) that was fixed at 7 days post-virus (DPV) and co-labeled with antibodies to BrdU (red) and Ki-67 (green). Phalloidin labeling (grayscale) is shown to aid in visualizing the borders of the sensory epithelium. White dashed lines demarcate the borders of the sensory epithelium. Scale bar, 100 µm. Insets show high-resolution views of nuclei in the sensory epithelium. Arrows indicate nuclei that labeled with antibodies to BrdU but not Ki67. Scale bar for insets, 5 µm. ( B ) Graph shows the mean number of BrdU-labeled nuclei (green data points) and Ki-67-labeled nuclei (red data points) per sensory epithelium at 5, 7, and 10 DPV. ( C ) Graph shows quantification of the percentage of the BrdU-positive population that did not label with Ki67 antibodies (green data points) and the percentage of the Ki-67-positive population that did not label with BrdU antibodies (red data points). ( D ) Confocal image of an adult mouse utricle infected with Ad.MT58A (1×10 9 TU/mL) that was fixed at 7 DPV and co-labeled with antibodies to PH3-Ser10 (white) and Ki-67 (red). ( E ) Confocal images of a utricle from an embryonic day 17.5 (E17.5) mouse that was fixed in vivo and co-labeled with antibodies to PH3-Ser10 (white) and Ki-67 (red). Phalloidin labeling (grayscale) is shown to aid in visualizing the borders of the sensory epithelium. White dashed lines demarcate the borders of the sensory epithelium, and arrows in D and E indicate PH3-Ser10/Ki-67 co-labeled nuclei. Scale bar for D–E, 100 µm. ( F ) Graph shows quantification of the percentage of the Ki-67-positive populations that labeled with antibodies to PH3-Ser10. The difference in the percentage of PH3-Ser10-positive/Ki-67-positive nuclei did not reach statistical significance (p > 0.05; Student's t-test). The numbers above the gray bars indicate the mean number of PH3-Ser10-labeled nuclei per sensory epithelium. There were significantly more PH3-Ser10-positive nuclei in E17.5 utricles (p

    Techniques Used: Infection, Labeling, In Vivo

    5) Product Images from "BMP signaling regulates satellite cell-dependent postnatal muscle growth"

    Article Title: BMP signaling regulates satellite cell-dependent postnatal muscle growth

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.144089

    Effects of Nog-mediated abrogation of BMP signaling on postnatal satellite cell activity. The anterior compartment of the lower hindlimb was transduced with AAV- Nog at P3. At P14, the mice were treated for 3 consecutive days with subcutaneous injections of BrdU and were sacrificed at P17. (A) Fluorescence images of a mid-belly transverse section of a TA muscle following immunostaining against Pax7 in red (arrows), against BrdU (green) and against laminin (white); nuclei were stained with DAPI (blue). Pax7 + /BrdU + cells are indicated with white arrows. Pax7 + /BrdU − satellite cells and Pax7 − /BrdU + sublaminal myonuclei are, respectively, indicated with red and green arrows. Scale bar: 20 µm. (B) The diagrams depict the number of Pax7 + , Pax7 + BrdU − and Pax7 + BrdU + satellite cells, as well as the number of newly recruited Pax7 − BrdU + myonuclei per 100 myofibers from the TA muscle. n =4 biological replicates for each condition. (C,D) The anterior compartment of the lower hindlimb was transduced with AAV- Nog at P3. Muscles were analyzed at 4 weeks of age. The diagram depicts the number of Pax7 + (C) and of m-Cadherin + (D) satellite cells per 100 myofibers quantified from whole TA muscle sections ( n =3 or 4 for saline-injected control; n =3 for AAV- Nog -injected muscle). Horizontal lines represent the median and dots represent individual data points.
    Figure Legend Snippet: Effects of Nog-mediated abrogation of BMP signaling on postnatal satellite cell activity. The anterior compartment of the lower hindlimb was transduced with AAV- Nog at P3. At P14, the mice were treated for 3 consecutive days with subcutaneous injections of BrdU and were sacrificed at P17. (A) Fluorescence images of a mid-belly transverse section of a TA muscle following immunostaining against Pax7 in red (arrows), against BrdU (green) and against laminin (white); nuclei were stained with DAPI (blue). Pax7 + /BrdU + cells are indicated with white arrows. Pax7 + /BrdU − satellite cells and Pax7 − /BrdU + sublaminal myonuclei are, respectively, indicated with red and green arrows. Scale bar: 20 µm. (B) The diagrams depict the number of Pax7 + , Pax7 + BrdU − and Pax7 + BrdU + satellite cells, as well as the number of newly recruited Pax7 − BrdU + myonuclei per 100 myofibers from the TA muscle. n =4 biological replicates for each condition. (C,D) The anterior compartment of the lower hindlimb was transduced with AAV- Nog at P3. Muscles were analyzed at 4 weeks of age. The diagram depicts the number of Pax7 + (C) and of m-Cadherin + (D) satellite cells per 100 myofibers quantified from whole TA muscle sections ( n =3 or 4 for saline-injected control; n =3 for AAV- Nog -injected muscle). Horizontal lines represent the median and dots represent individual data points.

    Techniques Used: Activity Assay, Transduction, Mouse Assay, Fluorescence, Immunostaining, Staining, Injection

    6) Product Images from "Selective proliferative response of microglia to alternative polarization signals"

    Article Title: Selective proliferative response of microglia to alternative polarization signals

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-017-1011-6

    Proliferation of peritoneal macrophages following IL-4 treatment in vivo . Ki67 protein expression ( a , b ) and BrdU incorporation ( c , d ) were analyzed in peritoneal macrophages isolated from mice treated i.p . with vehicle (veh) or 5 μg IL-4 for 24 h. a , c show representative dot plots depicting gating schemes for Ki67 and BrdU analyses, respectively. Bar charts represent the percentage of macrophages showing a positive signal for Ki67 ( b ) or BrdU ( d ) with respect to the total number of macrophages obtained from each mouse following the specified treatment. Bars represent the mean ± SEM of six mice per group. Student’s unpaired t test, *** p
    Figure Legend Snippet: Proliferation of peritoneal macrophages following IL-4 treatment in vivo . Ki67 protein expression ( a , b ) and BrdU incorporation ( c , d ) were analyzed in peritoneal macrophages isolated from mice treated i.p . with vehicle (veh) or 5 μg IL-4 for 24 h. a , c show representative dot plots depicting gating schemes for Ki67 and BrdU analyses, respectively. Bar charts represent the percentage of macrophages showing a positive signal for Ki67 ( b ) or BrdU ( d ) with respect to the total number of macrophages obtained from each mouse following the specified treatment. Bars represent the mean ± SEM of six mice per group. Student’s unpaired t test, *** p

    Techniques Used: In Vivo, Expressing, BrdU Incorporation Assay, Isolation, Mouse Assay

    Related Articles

    Labeling:

    Article Title: SIRT1 Deacetylates TopBP1 and Modulates Intra-S-Phase Checkpoint and DNA Replication Origin Firing
    Article Snippet: BrdU incorporation assay MEF cells were cultured overnight in 10 mm dishes. .. The cells were then either directly labeled with BrdU (BD), or treated with 10 mM hydroxyurea (HU) for 4 hours and released into medium containing BrdU, or reverse transfected by LipofectamineTM 2000 (Invitrogen) and then followed with the indicated treatment. .. The detailed FACS analysis of BrdU-containing samples was performed as described .

    Transfection:

    Article Title: SIRT1 Deacetylates TopBP1 and Modulates Intra-S-Phase Checkpoint and DNA Replication Origin Firing
    Article Snippet: BrdU incorporation assay MEF cells were cultured overnight in 10 mm dishes. .. The cells were then either directly labeled with BrdU (BD), or treated with 10 mM hydroxyurea (HU) for 4 hours and released into medium containing BrdU, or reverse transfected by LipofectamineTM 2000 (Invitrogen) and then followed with the indicated treatment. .. The detailed FACS analysis of BrdU-containing samples was performed as described .

    Chromatin Immunoprecipitation:

    Article Title: Arabidopsis thaliana Chromosome 4 Replicates in Two Phases That Correlate with Chromatin State
    Article Snippet: Genomic DNA extracted from the sorted nuclei was sonicated in 450 µL of ChIP dilution buffer (0.1% BSA, 1.2 mM EDTA, 16.7 mM Tris-HCl pH 8, and 167 mM NaCl) to a shear-size of 500 to 1000 bp, followed by addition of Triton X-100 (1.1%). .. The sheared DNA was denatured at 95°C for 5 min and immediately cooled on ice for at least 5 min. One mL of cold ChIP dilution buffer containing 1.1% Triton X-100 was added and the sheared DNA was incubated with 0.5 µL anti-BrdU antibody (Invitrogen) for 3 h at 4°C. .. DNA containing BrdU was immunoprecipitated by adding 100 µL of 50% protein G-sepharose beads (Sigma) and incubating overnight in the dark at 4°C with gentle agitation.

    Incubation:

    Article Title: Arabidopsis thaliana Chromosome 4 Replicates in Two Phases That Correlate with Chromatin State
    Article Snippet: Genomic DNA extracted from the sorted nuclei was sonicated in 450 µL of ChIP dilution buffer (0.1% BSA, 1.2 mM EDTA, 16.7 mM Tris-HCl pH 8, and 167 mM NaCl) to a shear-size of 500 to 1000 bp, followed by addition of Triton X-100 (1.1%). .. The sheared DNA was denatured at 95°C for 5 min and immediately cooled on ice for at least 5 min. One mL of cold ChIP dilution buffer containing 1.1% Triton X-100 was added and the sheared DNA was incubated with 0.5 µL anti-BrdU antibody (Invitrogen) for 3 h at 4°C. .. DNA containing BrdU was immunoprecipitated by adding 100 µL of 50% protein G-sepharose beads (Sigma) and incubating overnight in the dark at 4°C with gentle agitation.

    Mouse Assay:

    Article Title: p38 MAPK regulates Bax activity and apoptosis in enterocytes at baseline and after intestinal resection
    Article Snippet: .. Ninety minutes before death, mice were given an intraperitoneal injection of 5-bromodeoxyuridine (BrdU; 0.1 ml/10 gm body wt; Zymed Laboratories, San Francisco, CA). ..

    Article Title: Fast Renewal of the Distal Colonic Mucus Layers by the Surface Goblet Cells as Measured by In Vivo Labeling of Mucin Glycoproteins
    Article Snippet: .. Intraperitoneal Injection of GalNAz and BrdU in Mice and Tissue Fixation The Click iT reagent GalNAz (Invitrogen) was dissolved in 25 µl DMSO and diluted in PBS to a final volume of 500 µl per 2.6 mg reagent for each animal. .. The solution was supplemented with 3 mg 5-bromo-2'-deoxyuridine (BrdU, Invitrogen) per animal.

    Injection:

    Article Title: p38 MAPK regulates Bax activity and apoptosis in enterocytes at baseline and after intestinal resection
    Article Snippet: .. Ninety minutes before death, mice were given an intraperitoneal injection of 5-bromodeoxyuridine (BrdU; 0.1 ml/10 gm body wt; Zymed Laboratories, San Francisco, CA). ..

    Article Title: Fast Renewal of the Distal Colonic Mucus Layers by the Surface Goblet Cells as Measured by In Vivo Labeling of Mucin Glycoproteins
    Article Snippet: .. Intraperitoneal Injection of GalNAz and BrdU in Mice and Tissue Fixation The Click iT reagent GalNAz (Invitrogen) was dissolved in 25 µl DMSO and diluted in PBS to a final volume of 500 µl per 2.6 mg reagent for each animal. .. The solution was supplemented with 3 mg 5-bromo-2'-deoxyuridine (BrdU, Invitrogen) per animal.

    Immunolabeling:

    Article Title: Depletion of nuclear actin is a key mediator of quiescence in epithelial cells
    Article Snippet: YFP–LAP was generated from CMV-LAP, which was kindly provided by Ueli Schibler (University of Geneva, Switzerland). .. The following antibodies were used: rabbit-anti-lamin A/C (Santa Cruz 1/500), -anti-β-actin (ab1801 from Abcam; used for immunolabeling experiments at 1/100), -anti-histone H3 (Abcam, 1/10,000), -anti-acetylated histone H4 (Upstate Biotech, 1/500), -anti-cofilin (Abcam, 1/1000), -anti-Pol II (Santa Cruz: N20, 1/100), -anti-RNA Pol II P -Ser5 (Abcam, 1/500), -anti-RPC32 (Santa Cruz Biotech, CA), -anti-FLAG (Sigma, 1/750); mouse-anti-β-actin [A5441 (Clone AC-15) from Sigma, used for western blot analysis at 1/1000], -anti-β-actin [Ab3280 (ACTN05-C4) from Abcam, used for western blot analysis at 1:1000], -anti-bromodeoxyuridine (BrdU) (Invitrogen, 1/75; Roche, 1/10), -anti-FLAG (Sigma, 1/750); rat-anti-BrdU (Abcam, 1/100), -anti-LN1 α1 chain (clones 198 and 200; a kind gift from Lydia Sorokin, 1/50); sheep anti-BrdU (Abcam, 1/100); anti-rabbit, -mouse, -rat and -sheep secondary antibodies conjugated to Alexa Fluor 488 or 594 (Invitrogen, 1/400). .. Finally, DNase I conjugated to Alexa Fluor 594 (DNase I-594: Invitrogen, 1/500) was used.

    Western Blot:

    Article Title: Depletion of nuclear actin is a key mediator of quiescence in epithelial cells
    Article Snippet: YFP–LAP was generated from CMV-LAP, which was kindly provided by Ueli Schibler (University of Geneva, Switzerland). .. The following antibodies were used: rabbit-anti-lamin A/C (Santa Cruz 1/500), -anti-β-actin (ab1801 from Abcam; used for immunolabeling experiments at 1/100), -anti-histone H3 (Abcam, 1/10,000), -anti-acetylated histone H4 (Upstate Biotech, 1/500), -anti-cofilin (Abcam, 1/1000), -anti-Pol II (Santa Cruz: N20, 1/100), -anti-RNA Pol II P -Ser5 (Abcam, 1/500), -anti-RPC32 (Santa Cruz Biotech, CA), -anti-FLAG (Sigma, 1/750); mouse-anti-β-actin [A5441 (Clone AC-15) from Sigma, used for western blot analysis at 1/1000], -anti-β-actin [Ab3280 (ACTN05-C4) from Abcam, used for western blot analysis at 1:1000], -anti-bromodeoxyuridine (BrdU) (Invitrogen, 1/75; Roche, 1/10), -anti-FLAG (Sigma, 1/750); rat-anti-BrdU (Abcam, 1/100), -anti-LN1 α1 chain (clones 198 and 200; a kind gift from Lydia Sorokin, 1/50); sheep anti-BrdU (Abcam, 1/100); anti-rabbit, -mouse, -rat and -sheep secondary antibodies conjugated to Alexa Fluor 488 or 594 (Invitrogen, 1/400). .. Finally, DNase I conjugated to Alexa Fluor 594 (DNase I-594: Invitrogen, 1/500) was used.

    Flow Cytometry:

    Article Title: TGF-? Signalling Is Required for CD4+ T Cell Homeostasis But Dispensable for Regulatory T Cell Function
    Article Snippet: For adoptive transfer experiments, Rag−/− mice were injected intraperitonelly with 2×106 purified T cells (T cell isolation kit, Milteny). .. Flow Cytometric Analyses For flow cytometry the following fluorochrome conjugated antibodies were used: anti-CD3, anti-CD4 (L3T4), anti-CD8α (53-6.7), anti-CD8β (YTS 156.7.7), anti-CD19, anti-CD25, anti-CD44, anti-CD45, anti-CD45.1, anti-CD45.2, anti-CD45RB, anti-CD49b, anti-CD62L, anti-CD69, anti-CD122, anti-CD154, anti-GITR, anti-NK1.1, anti-CCR4, anti-CCR5, anti-CCR6, anti-CCR7, anti-IL2, anti-Ki-67, anti-INF-γ, anti-IL-4 all from BD Biosciences, anti-TGF-βRII from R & D, anti-Ly6A/E, anti-CD90.2 from BioLegend anti-Foxp3, and anti-BrdU from eBiosciences from Cell Signalling Technology. .. Staining for Foxp3, BrdU, and Ki67 was performed using Foxp3 Staining Buffer Set (eBiosciences) according to the manufacturer's protocol.

    Cytometry:

    Article Title: TGF-? Signalling Is Required for CD4+ T Cell Homeostasis But Dispensable for Regulatory T Cell Function
    Article Snippet: For adoptive transfer experiments, Rag−/− mice were injected intraperitonelly with 2×106 purified T cells (T cell isolation kit, Milteny). .. Flow Cytometric Analyses For flow cytometry the following fluorochrome conjugated antibodies were used: anti-CD3, anti-CD4 (L3T4), anti-CD8α (53-6.7), anti-CD8β (YTS 156.7.7), anti-CD19, anti-CD25, anti-CD44, anti-CD45, anti-CD45.1, anti-CD45.2, anti-CD45RB, anti-CD49b, anti-CD62L, anti-CD69, anti-CD122, anti-CD154, anti-GITR, anti-NK1.1, anti-CCR4, anti-CCR5, anti-CCR6, anti-CCR7, anti-IL2, anti-Ki-67, anti-INF-γ, anti-IL-4 all from BD Biosciences, anti-TGF-βRII from R & D, anti-Ly6A/E, anti-CD90.2 from BioLegend anti-Foxp3, and anti-BrdU from eBiosciences from Cell Signalling Technology. .. Staining for Foxp3, BrdU, and Ki67 was performed using Foxp3 Staining Buffer Set (eBiosciences) according to the manufacturer's protocol.

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    Thermo Fisher brdu
    <t>BrdU</t> labeling of transplanted SC is shown. SC were transplanted in syngeneic rats ( A and B ) and immunocompromised mice ( C and D ). Tissue sections were double-immunostained for <t>WT1</t> (green, A – D ) and BrdU (red, A – D ). B and D ) Higher magnification
    Brdu, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94
    Thermo Fisher brdu labeling
    1,25(OH) 2 D 3 maintains <t>enterocyte</t> turnover in CCl 4 -treated rats. (A) <t>BrdU</t> expression in small intestine 6 h after BrdU injection. (B) Quantification of BrdU+ cells along the crypt-villus axis. * p
    Brdu Labeling, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BrdU labeling of transplanted SC is shown. SC were transplanted in syngeneic rats ( A and B ) and immunocompromised mice ( C and D ). Tissue sections were double-immunostained for WT1 (green, A – D ) and BrdU (red, A – D ). B and D ) Higher magnification

    Journal: Biology of Reproduction

    Article Title: Nondividing, Postpubertal Rat Sertoli Cells Resumed Proliferation after Transplantation 1

    doi: 10.1095/biolreprod.113.110197

    Figure Lengend Snippet: BrdU labeling of transplanted SC is shown. SC were transplanted in syngeneic rats ( A and B ) and immunocompromised mice ( C and D ). Tissue sections were double-immunostained for WT1 (green, A – D ) and BrdU (red, A – D ). B and D ) Higher magnification

    Article Snippet: To detect BrdU-labeled proliferating SC, double- immunofluorescence staining was performed for WT1 and BrdU, using Alexa Fluor 488-labeled anti-mouse (1:400 dilution; Invitrogen) and Alexa Fluor 594 anti-rabbit (1:400 dilution; Invitrogen) secondary antibodies.

    Techniques: Labeling, Mouse Assay

    Flow cytometric analysis of Arabidopsis cell suspension culture. (A) Analytical FACS profile showing BrdU incorporation into Arabidopsis nuclei as a function of DNA content. BrdU incorporation and DNA content of nuclei were visualized using anti-BrdU Alexa 488 conjugate and propidium iodide, respectively. Five boxes are shown, representing nuclei in G1, early S, mid S, late S, and G2/M, respectively. (B) Histogram plots for total and BrdU-positive nuclei from the BrdU labeled cells shown in (A). (C) Flow diagram of FACS-based microarray experiments for profiling replication in early, mid and late S. Cells were pulse-labeled with BrdU, and nuclei isolated. Populations of nuclei in early S/G1, mid S, and late S/G2 were sorted based on DNA content. Genomic DNA was prepared from the sorted nuclei in each fraction and sheared to an average size of 500 bp before heat denaturation and immunoprecipitation with antibodies against BrdU. DNA containing BrdU was amplified, labeled with Cy dyes and hybridized to a tiling array for Arabidopsis chr 4.

    Journal: PLoS Genetics

    Article Title: Arabidopsis thaliana Chromosome 4 Replicates in Two Phases That Correlate with Chromatin State

    doi: 10.1371/journal.pgen.1000982

    Figure Lengend Snippet: Flow cytometric analysis of Arabidopsis cell suspension culture. (A) Analytical FACS profile showing BrdU incorporation into Arabidopsis nuclei as a function of DNA content. BrdU incorporation and DNA content of nuclei were visualized using anti-BrdU Alexa 488 conjugate and propidium iodide, respectively. Five boxes are shown, representing nuclei in G1, early S, mid S, late S, and G2/M, respectively. (B) Histogram plots for total and BrdU-positive nuclei from the BrdU labeled cells shown in (A). (C) Flow diagram of FACS-based microarray experiments for profiling replication in early, mid and late S. Cells were pulse-labeled with BrdU, and nuclei isolated. Populations of nuclei in early S/G1, mid S, and late S/G2 were sorted based on DNA content. Genomic DNA was prepared from the sorted nuclei in each fraction and sheared to an average size of 500 bp before heat denaturation and immunoprecipitation with antibodies against BrdU. DNA containing BrdU was amplified, labeled with Cy dyes and hybridized to a tiling array for Arabidopsis chr 4.

    Article Snippet: The sheared DNA was denatured at 95°C for 5 min and immediately cooled on ice for at least 5 min. One mL of cold ChIP dilution buffer containing 1.1% Triton X-100 was added and the sheared DNA was incubated with 0.5 µL anti-BrdU antibody (Invitrogen) for 3 h at 4°C.

    Techniques: Flow Cytometry, FACS, BrdU Incorporation Assay, Labeling, Microarray, Isolation, Immunoprecipitation, Amplification

    Arabidopsis chromosome 4 with replication profiles for early, mid, and late S phase suspension culture cells. (A) Gene and TE coverage were determined using TAIR8 annotation and are expressed as percentage of bases in 1 kb non-overlapping segments occurring in genes or TEs respectively. Overlapping genes or TEs were merged to prevent coverage values exceeding 100%. Data were loess-smoothed using a 150 kb window. (B) GC percentage calculated in 1 kb non-overlapping windows and loess-smoothed using a 150 kb window. (C) Schematic representation of chromosome 4 omitting the telomeres and nucleolar organizing region. The gene-rich euchromatic distal short and distal long arms are shaded light gray while the heterochromatic knob and pericentromere are rich in TEs and are shaded black. The proximal portions of both the short and long arms have intermediate characteristics and are shaded dark gray. (D) Replication profiles for early, mid, and late S phase cells. Replication is expressed as log 2 ratio of BrdU-labeled sequences in early, mid or late S phase cells with respect to total DNA from the same cells. Data have been normalized and scaled within experiments, but no normalization was performed between experiments.

    Journal: PLoS Genetics

    Article Title: Arabidopsis thaliana Chromosome 4 Replicates in Two Phases That Correlate with Chromatin State

    doi: 10.1371/journal.pgen.1000982

    Figure Lengend Snippet: Arabidopsis chromosome 4 with replication profiles for early, mid, and late S phase suspension culture cells. (A) Gene and TE coverage were determined using TAIR8 annotation and are expressed as percentage of bases in 1 kb non-overlapping segments occurring in genes or TEs respectively. Overlapping genes or TEs were merged to prevent coverage values exceeding 100%. Data were loess-smoothed using a 150 kb window. (B) GC percentage calculated in 1 kb non-overlapping windows and loess-smoothed using a 150 kb window. (C) Schematic representation of chromosome 4 omitting the telomeres and nucleolar organizing region. The gene-rich euchromatic distal short and distal long arms are shaded light gray while the heterochromatic knob and pericentromere are rich in TEs and are shaded black. The proximal portions of both the short and long arms have intermediate characteristics and are shaded dark gray. (D) Replication profiles for early, mid, and late S phase cells. Replication is expressed as log 2 ratio of BrdU-labeled sequences in early, mid or late S phase cells with respect to total DNA from the same cells. Data have been normalized and scaled within experiments, but no normalization was performed between experiments.

    Article Snippet: The sheared DNA was denatured at 95°C for 5 min and immediately cooled on ice for at least 5 min. One mL of cold ChIP dilution buffer containing 1.1% Triton X-100 was added and the sheared DNA was incubated with 0.5 µL anti-BrdU antibody (Invitrogen) for 3 h at 4°C.

    Techniques: Labeling

    1,25(OH) 2 D 3 maintains enterocyte turnover in CCl 4 -treated rats. (A) BrdU expression in small intestine 6 h after BrdU injection. (B) Quantification of BrdU+ cells along the crypt-villus axis. * p

    Journal: Biomolecules & Therapeutics

    Article Title: Vitamin D Improves Intestinal Barrier Function in Cirrhosis Rats by Upregulating Heme Oxygenase-1 Expression

    doi: 10.4062/biomolther.2018.052

    Figure Lengend Snippet: 1,25(OH) 2 D 3 maintains enterocyte turnover in CCl 4 -treated rats. (A) BrdU expression in small intestine 6 h after BrdU injection. (B) Quantification of BrdU+ cells along the crypt-villus axis. * p

    Article Snippet: BrdU labeling and enterocyte migration Standard 5-bromodeoxyuridine (BrdU) labeling reagent (Zymed Laboratories) was injected intraperitoneally at a dose of 50 mg/kg body weight 6 h before the animals were euthanized.

    Techniques: Expressing, Injection

    Effect of acetylation inhibition by C-646 on the ability of IFI16 to recognize and bind the KSHV genome. (A and B) HMVEC-d cells pre-incubated with or without 1 μM C-646 for 2 h were washed, infected with BrdU genome labeled KSHV (30 DNA copies/cell) for 2 h, washed and incubated with or without 1 μM C-646 for 4 h. Cells were processed, incubated with anti-BrdU antibodies, washed, and reacted with Alexa Fluor-488 (green) secondary antibodies to detect BrdU-KSHV. These slides were subjected to PLA (A) with mouse anti-IFI16 and rabbit-IFI16 antibodies or (B) with anti-IFI16 and anti-acetylated lysine antibodies. Boxed areas are enlarged. The yellow arrows in (A) and (B) indicate the cytoplasmic IFI16 or acetylated IFI16, respectively. The white arrows in (A) indicate colocalization of IFI16 PLA spots with BrdU labeled KSHV genome. The white arrows in (B) indicate colocalization of acetylated IFI16 PLA spots with BrdU labeled KSHV genome. (C and D) Bar graph of quantitation of colocalization of IFI16 or acetylated IFI16 PLA spots with BrdU labeled KSHV genome in (A) and (B) panels. At least ten fields with a minimum of 3–4 cells/field were counted. (E) The above described cells were subjected to PLA using anti-BrdU and anti-IFI16 antibodies to detect the direct association of IFI16 with BrdU-labeled KSHV genome. Boxed areas are enlarged. The PLA dots indicated by white arrows represent the association between IFI16 and BrdU-labeled KSHV genome. (F) PLA spots from 10 fields with at least 3–4 cells/field in (E) were quantitated and presented as a bar graph (G) Chromatin immunoprecipitation was performed using anti-IFI16 antibody as described in the Material and Methods. Untreated BCBL-1 cells (3 x 10 7 ) or cells treated with 1 μM C-646 for 24 h were cross-linked with formaldehyde and sonicated to obtain DNA fragments between 200–400 bps. q-PCR was performed with primers for two different regions on the KSHV genome and one control for GAPDH, and the results are presented as fold enrichment of immunoprecipitated DNA. The p values were calculated using Student’s T test. NS (non-significant).

    Journal: PLoS Pathogens

    Article Title: Herpesvirus Genome Recognition Induced Acetylation of Nuclear IFI16 Is Essential for Its Cytoplasmic Translocation, Inflammasome and IFN-β Responses

    doi: 10.1371/journal.ppat.1005019

    Figure Lengend Snippet: Effect of acetylation inhibition by C-646 on the ability of IFI16 to recognize and bind the KSHV genome. (A and B) HMVEC-d cells pre-incubated with or without 1 μM C-646 for 2 h were washed, infected with BrdU genome labeled KSHV (30 DNA copies/cell) for 2 h, washed and incubated with or without 1 μM C-646 for 4 h. Cells were processed, incubated with anti-BrdU antibodies, washed, and reacted with Alexa Fluor-488 (green) secondary antibodies to detect BrdU-KSHV. These slides were subjected to PLA (A) with mouse anti-IFI16 and rabbit-IFI16 antibodies or (B) with anti-IFI16 and anti-acetylated lysine antibodies. Boxed areas are enlarged. The yellow arrows in (A) and (B) indicate the cytoplasmic IFI16 or acetylated IFI16, respectively. The white arrows in (A) indicate colocalization of IFI16 PLA spots with BrdU labeled KSHV genome. The white arrows in (B) indicate colocalization of acetylated IFI16 PLA spots with BrdU labeled KSHV genome. (C and D) Bar graph of quantitation of colocalization of IFI16 or acetylated IFI16 PLA spots with BrdU labeled KSHV genome in (A) and (B) panels. At least ten fields with a minimum of 3–4 cells/field were counted. (E) The above described cells were subjected to PLA using anti-BrdU and anti-IFI16 antibodies to detect the direct association of IFI16 with BrdU-labeled KSHV genome. Boxed areas are enlarged. The PLA dots indicated by white arrows represent the association between IFI16 and BrdU-labeled KSHV genome. (F) PLA spots from 10 fields with at least 3–4 cells/field in (E) were quantitated and presented as a bar graph (G) Chromatin immunoprecipitation was performed using anti-IFI16 antibody as described in the Material and Methods. Untreated BCBL-1 cells (3 x 10 7 ) or cells treated with 1 μM C-646 for 24 h were cross-linked with formaldehyde and sonicated to obtain DNA fragments between 200–400 bps. q-PCR was performed with primers for two different regions on the KSHV genome and one control for GAPDH, and the results are presented as fold enrichment of immunoprecipitated DNA. The p values were calculated using Student’s T test. NS (non-significant).

    Article Snippet: For generating 5-bromo-2-deoxyuridine (BrdU) incorporated KSHV genome, BrdU labeling reagent (Life Technologies) was added to the culture medium in a 1:100 (v:v) ratio (from the supplied stock) [ ].

    Techniques: Inhibition, Incubation, Infection, Labeling, Proximity Ligation Assay, Quantitation Assay, Chromatin Immunoprecipitation, Sonication, Polymerase Chain Reaction, Immunoprecipitation

    Colocalization of IFI16 with BrdU genome labeled KSHV in the nucleus, acetylation of IFI16 in the nucleus and cytoplasmic redistribution during de novo KSHV infection of HMVEC-d cells. (A) HMVEC-d cells were infected for 15 and 30 min with BrdU genome labeled KSHV (30 DNA copies/cell), processed for IFA, reacted with anti-IFI16 and anti-BrdU antibodies followed by Alexa Fluor-594/488 secondary antibodies and DAPI (blue). The boxed areas are enlarged. Red arrows: KSHV genome in the cytoplasm (green dots); yellow arrows: cytoplasmic IFI16; white arrows: colocalization of IFI16 with KSHV genome (yellow spots) in the nucleus. 60X magnification. (B) HMVEC-d cells were uninfected (UI) or infected with KSHV for different time points, nuclear and cytoplasmic fractions isolated and western blotted for IFI16. Tubulin and TBP were used as purity markers for cytoplasmic and nuclear fractions, respectively, and as loading controls. (C and D) Equal quantities of whole cell lysate (WCL) proteins from uninfected and KSHV infected (24 h) cells in NETN buffer were immunoprecipitated (IP-ed) with anti-acetylated lysine antibody and western blotted for IFI16 and tubulin (C), and cytoplasmic and nuclear proteins (D) were IP-ed with anti-acetylated lysine antibody and western blotted for IFI16, tubulin and H3. Cytoplasmic and nuclear proteins were western blotted for total H3, tubulin and TBP. (E) HMVEC-d cells were serum-starved without (UT = untreated) or with 1μM p300 inhibitor C-646 for 2 h, uninfected or infected with KSHV for 2 h, washed and incubated in complete medium in the presence or absence of inhibitor for 4 and 24 h. WCL in NETN buffer were IP-ed with anti-acetylated lysine or IFI16 antibodies and western blotted for IFI16. (F) 24 h lysates from (E) were IP-ed with anti-IFI16 and western blotted for acetylated lysine. (G) HMVEC-d cells treated or untreated with C-646 for 2 h were either left uninfected (UI) or infected with KSHV for various time points, nuclear and cytoplasmic fractions isolated and western blotted for IFI16.

    Journal: PLoS Pathogens

    Article Title: Herpesvirus Genome Recognition Induced Acetylation of Nuclear IFI16 Is Essential for Its Cytoplasmic Translocation, Inflammasome and IFN-β Responses

    doi: 10.1371/journal.ppat.1005019

    Figure Lengend Snippet: Colocalization of IFI16 with BrdU genome labeled KSHV in the nucleus, acetylation of IFI16 in the nucleus and cytoplasmic redistribution during de novo KSHV infection of HMVEC-d cells. (A) HMVEC-d cells were infected for 15 and 30 min with BrdU genome labeled KSHV (30 DNA copies/cell), processed for IFA, reacted with anti-IFI16 and anti-BrdU antibodies followed by Alexa Fluor-594/488 secondary antibodies and DAPI (blue). The boxed areas are enlarged. Red arrows: KSHV genome in the cytoplasm (green dots); yellow arrows: cytoplasmic IFI16; white arrows: colocalization of IFI16 with KSHV genome (yellow spots) in the nucleus. 60X magnification. (B) HMVEC-d cells were uninfected (UI) or infected with KSHV for different time points, nuclear and cytoplasmic fractions isolated and western blotted for IFI16. Tubulin and TBP were used as purity markers for cytoplasmic and nuclear fractions, respectively, and as loading controls. (C and D) Equal quantities of whole cell lysate (WCL) proteins from uninfected and KSHV infected (24 h) cells in NETN buffer were immunoprecipitated (IP-ed) with anti-acetylated lysine antibody and western blotted for IFI16 and tubulin (C), and cytoplasmic and nuclear proteins (D) were IP-ed with anti-acetylated lysine antibody and western blotted for IFI16, tubulin and H3. Cytoplasmic and nuclear proteins were western blotted for total H3, tubulin and TBP. (E) HMVEC-d cells were serum-starved without (UT = untreated) or with 1μM p300 inhibitor C-646 for 2 h, uninfected or infected with KSHV for 2 h, washed and incubated in complete medium in the presence or absence of inhibitor for 4 and 24 h. WCL in NETN buffer were IP-ed with anti-acetylated lysine or IFI16 antibodies and western blotted for IFI16. (F) 24 h lysates from (E) were IP-ed with anti-IFI16 and western blotted for acetylated lysine. (G) HMVEC-d cells treated or untreated with C-646 for 2 h were either left uninfected (UI) or infected with KSHV for various time points, nuclear and cytoplasmic fractions isolated and western blotted for IFI16.

    Article Snippet: For generating 5-bromo-2-deoxyuridine (BrdU) incorporated KSHV genome, BrdU labeling reagent (Life Technologies) was added to the culture medium in a 1:100 (v:v) ratio (from the supplied stock) [ ].

    Techniques: Labeling, Infection, Immunofluorescence, Isolation, Western Blot, Immunoprecipitation, Incubation