in situ brdu red dna fragmentation tunel assay kit  (Abcam)

 
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    In situ BrdU Red DNA Fragmentation TUNEL Assay Kit
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    ab66110
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    Structured Review

    Abcam in situ brdu red dna fragmentation tunel assay kit
    <t>TUNEL-positive</t> cells increased after HEV infection. TUNEL staining of the HEV-RNA positive brain tissues collected on 14, 21, and 28 dpi was performed using in situ <t>BrdU-Red</t> <t>DNA</t> Fragmentation (TUNEL) Assay Kit and slides were counterstained with DAPI (Blue). TUNEL-positive signals merged with DAPI were shown as purple. (A,B) Few positive signals were detected in animals of mock group. (C–E) Positive signals were exhibited in infected brain sections. The high magnification images (D,E) depicted granular masses of positive signals at nucleus of the apoptotic cells. (F) TUNEL-positive cells were significantly increased in HEV-infected gerbil brain tissues (* p

    https://www.bioz.com/result/in situ brdu red dna fragmentation tunel assay kit/product/Abcam
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    in situ brdu red dna fragmentation tunel assay kit - by Bioz Stars, 2021-03
    95/100 stars

    Images

    1) Product Images from "Hepatitis E Virus Induces Brain Injury Probably Associated With Mitochondrial Apoptosis"

    Article Title: Hepatitis E Virus Induces Brain Injury Probably Associated With Mitochondrial Apoptosis

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2019.00433

    TUNEL-positive cells increased after HEV infection. TUNEL staining of the HEV-RNA positive brain tissues collected on 14, 21, and 28 dpi was performed using in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit and slides were counterstained with DAPI (Blue). TUNEL-positive signals merged with DAPI were shown as purple. (A,B) Few positive signals were detected in animals of mock group. (C–E) Positive signals were exhibited in infected brain sections. The high magnification images (D,E) depicted granular masses of positive signals at nucleus of the apoptotic cells. (F) TUNEL-positive cells were significantly increased in HEV-infected gerbil brain tissues (* p
    Figure Legend Snippet: TUNEL-positive cells increased after HEV infection. TUNEL staining of the HEV-RNA positive brain tissues collected on 14, 21, and 28 dpi was performed using in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit and slides were counterstained with DAPI (Blue). TUNEL-positive signals merged with DAPI were shown as purple. (A,B) Few positive signals were detected in animals of mock group. (C–E) Positive signals were exhibited in infected brain sections. The high magnification images (D,E) depicted granular masses of positive signals at nucleus of the apoptotic cells. (F) TUNEL-positive cells were significantly increased in HEV-infected gerbil brain tissues (* p

    Techniques Used: TUNEL Assay, Infection, Staining, In Situ

    2) Product Images from "An SR protein is essential for the recovery of malaria parasites from DNA damage and exposure to artemisinin"

    Article Title: An SR protein is essential for the recovery of malaria parasites from DNA damage and exposure to artemisinin

    Journal: bioRxiv

    doi: 10.1101/2021.01.11.426314

    Pf SR1 expression is required to overcome artesunate exposure. (A). Western blot analysis demonstrating the increase in γ-PfH2A levels following exposure of tightly synchronized ring stage parasites to 700 nM artesunate for different time periods. (B). DNA fragmentation imaging by TUNEL assay of parasites exposed to 700 nM artesunate for 15 minutes, demonstrating increased levels of DNA breaks. Quantification of the ratio of TUNEL positive and negative nuclei in each treatment is presented on the right (n=100). Scale bar is 5 μm. (C). Growth curves of tightly synchronized ring stage Pf SR1- glmS (left) and NF54 parasites (right), treated with 700 nM artesunate for 6h. GlcN was either not added (Art, ), added 72h prior to artesunate treatment (Art+GlcN, ), or added continuously throughout the entire recovery period (Art+GlcN cont. ). Parasites growing on regular media without any treatment (no treatment, ) and parasites treated only with GlcN (GlcN, ) are presented as controls. The results presented are the average parasitemia of 3 biological replicates measured daily. Error bar represents standard errors.
    Figure Legend Snippet: Pf SR1 expression is required to overcome artesunate exposure. (A). Western blot analysis demonstrating the increase in γ-PfH2A levels following exposure of tightly synchronized ring stage parasites to 700 nM artesunate for different time periods. (B). DNA fragmentation imaging by TUNEL assay of parasites exposed to 700 nM artesunate for 15 minutes, demonstrating increased levels of DNA breaks. Quantification of the ratio of TUNEL positive and negative nuclei in each treatment is presented on the right (n=100). Scale bar is 5 μm. (C). Growth curves of tightly synchronized ring stage Pf SR1- glmS (left) and NF54 parasites (right), treated with 700 nM artesunate for 6h. GlcN was either not added (Art, ), added 72h prior to artesunate treatment (Art+GlcN, ), or added continuously throughout the entire recovery period (Art+GlcN cont. ). Parasites growing on regular media without any treatment (no treatment, ) and parasites treated only with GlcN (GlcN, ) are presented as controls. The results presented are the average parasitemia of 3 biological replicates measured daily. Error bar represents standard errors.

    Techniques Used: Expressing, Western Blot, Imaging, TUNEL Assay

    Pf SR1 is essential for parasite recovery from DNA damage cause by X-ray irradiation. (A). Exposure of Pf SR1- glmS parasite lines to increasing levels of X-ray irradiation is associated with increased levels of γ-PfH2A. (B). DNA fragmentation imaging by TUNEL assay of parasites exposed to increasing levels of X-ray irradiation demonstrating increased levels of DNA breaks. Quantification of the percentage of TUNEL positive and negative nuclei in each treatment is presented on the right (n=100). Scale bar is 5 μm. (C). Western blot analysis demonstrating that the levels of γ-PfH2A are elevated 15 minutes after X-ray irradiation regardless of GlcN treatment or parasite line (5mM for 72 h prior to irradiation). (D). Western blot analysis of Pf SR1- glmS parasite line growing on GlcN over time, indicating that γ-PfH2A accumulates approximately one week after Pf SR1 down-regulation. (E). Growth curves of wild type NF54 parasites exposed to a near lethal dose of X-ray irradiation of 6000 rad. Parasites were grown either on regular media (-GlcN) or with media added with 5mM GlcN for 72h and washed immediately before irradiation (+ GlcN). (F). Growth curves of Pf SR1- glmS parasites exposed to 6000 rad X-ray irradiation and grown either on regular media (-GlcN) or with media supplemented with 5mM GlcN for 72h and washed immediately before irradiation (+ GlcN). (G). Growth curves of wild type NF54 parasites exposed to 6000 rad X-ray irradiation and grown either on media supplemented continuously with 5mM GlcN (cont. GlcN) or on regular media. (H). Growth curves of Pf SR1- glmS parasites exposed to 6000 rad X-ray irradiation and grown either in media supplemented continuously with 5mM GlcN (cont. GlcN) or in regular media. Each of the curves represents the average parasitemia of 3 biological replicates at each timepoint. Error bars represent standard errors.
    Figure Legend Snippet: Pf SR1 is essential for parasite recovery from DNA damage cause by X-ray irradiation. (A). Exposure of Pf SR1- glmS parasite lines to increasing levels of X-ray irradiation is associated with increased levels of γ-PfH2A. (B). DNA fragmentation imaging by TUNEL assay of parasites exposed to increasing levels of X-ray irradiation demonstrating increased levels of DNA breaks. Quantification of the percentage of TUNEL positive and negative nuclei in each treatment is presented on the right (n=100). Scale bar is 5 μm. (C). Western blot analysis demonstrating that the levels of γ-PfH2A are elevated 15 minutes after X-ray irradiation regardless of GlcN treatment or parasite line (5mM for 72 h prior to irradiation). (D). Western blot analysis of Pf SR1- glmS parasite line growing on GlcN over time, indicating that γ-PfH2A accumulates approximately one week after Pf SR1 down-regulation. (E). Growth curves of wild type NF54 parasites exposed to a near lethal dose of X-ray irradiation of 6000 rad. Parasites were grown either on regular media (-GlcN) or with media added with 5mM GlcN for 72h and washed immediately before irradiation (+ GlcN). (F). Growth curves of Pf SR1- glmS parasites exposed to 6000 rad X-ray irradiation and grown either on regular media (-GlcN) or with media supplemented with 5mM GlcN for 72h and washed immediately before irradiation (+ GlcN). (G). Growth curves of wild type NF54 parasites exposed to 6000 rad X-ray irradiation and grown either on media supplemented continuously with 5mM GlcN (cont. GlcN) or on regular media. (H). Growth curves of Pf SR1- glmS parasites exposed to 6000 rad X-ray irradiation and grown either in media supplemented continuously with 5mM GlcN (cont. GlcN) or in regular media. Each of the curves represents the average parasitemia of 3 biological replicates at each timepoint. Error bars represent standard errors.

    Techniques Used: Irradiation, Imaging, TUNEL Assay, Western Blot

    3) Product Images from "Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments"

    Article Title: Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0225660

    More DNA fragmention of U-87MG and U-251 cells occurred after the combined SLCP and BBR therapy than by individual treatments. U-87MG and U-251MG cells were grown in EMEM and they were treated with either SLCP (20 μM), BBR (100 μM) or their combination (in the 1:5 ratio) for 48 h. The cells were stained with terminal deoxyribonucleic acid nick end labeling (TUNEL) reagents for detecting DNA fragmented cells and counter-stained with Hoechst 33342. A fluorescent microscope (Leica Germany) was used to detect the signal with appropriate excitation/emission filters. A-B: Representative images of TUNEL/Hoechst 33342 stained U-87MG (A) and U-251MG (B) cells after treatment with SLCP, BBR or their combination. C-D: More TUNEL-positive cells were observed in combination-treated cells in both GB cell lines than for individual treatments. Arrows indicate TUNEL positive cells. Scale bars indicate 100 μm and is applicable to all images. *p
    Figure Legend Snippet: More DNA fragmention of U-87MG and U-251 cells occurred after the combined SLCP and BBR therapy than by individual treatments. U-87MG and U-251MG cells were grown in EMEM and they were treated with either SLCP (20 μM), BBR (100 μM) or their combination (in the 1:5 ratio) for 48 h. The cells were stained with terminal deoxyribonucleic acid nick end labeling (TUNEL) reagents for detecting DNA fragmented cells and counter-stained with Hoechst 33342. A fluorescent microscope (Leica Germany) was used to detect the signal with appropriate excitation/emission filters. A-B: Representative images of TUNEL/Hoechst 33342 stained U-87MG (A) and U-251MG (B) cells after treatment with SLCP, BBR or their combination. C-D: More TUNEL-positive cells were observed in combination-treated cells in both GB cell lines than for individual treatments. Arrows indicate TUNEL positive cells. Scale bars indicate 100 μm and is applicable to all images. *p

    Techniques Used: Staining, End Labeling, TUNEL Assay, Microscopy

    Related Articles

    In Situ:

    Article Title: Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments
    Article Snippet: Chemicals Berberine (catalog no: B3251-10G), MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium], propidium iodide (PI), ethidium bromide (EtBr), agarose, proteinase-K protease inhibitor cocktail (catalog no: P8340-5ML) and other accessory chemicals were procured from Sigma (St. Louis, MO). .. An in situ BrdU-Red DNA fragmentation assay kit (TUNEL staining kit, catalog no: ab66110) and Annexin-V staining kit (catalog no: ab176749) were purchased from Abcam (Cambridge, MA). .. Low melting agarose was from Invitrogen (Grand Island, NY; catalog no: 16520050).

    Article Title: Class I HDAC inhibitor mocetinostat induces apoptosis by activation of miR-31 expression and suppression of E2F6
    Article Snippet: .. For in vivo apoptosis detection, the in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (Abcam, Cambridge, MA, USA) was used following the manufacturer’s protocol. ..

    Article Title: Mutant FUS causes DNA ligation defects to inhibit oxidative damage repair in Amyotrophic Lateral Sclerosis
    Article Snippet: TUNEL staining was used to analyze apoptosis in motor neurons. .. TUNEL Assay Kit-In situ BrdU-Red DNA Fragmentation (Abcam) NeuroTACS II in situ Apoptosis Detection kit (TREVIGEN) was used for the staining. ..

    Article Title: Inhibition of DYRK1A disrupts neural lineage specificationin human pluripotent stem cells
    Article Snippet: The cell cycle distribution of control and ID-8 treated pluripotent WA09 cells was analysed using the Click-IT EdU Flow Cytometry Kit AF488 (Invitrogen/Thermofisher, #C-10425). .. Apoptosis of control and ID-8 treated pluripotent WA09 cells was analysed using the Abcam In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (Abcam, Cambridge, MA). .. To identify gRNAs capable of driving CRISPRa activation of the DYRK1A gene, a number of unique 20mers in region between −250 to −50 bps relative to the main transcription start site were cloned in the pX462 gRNA and dCas9-VP64-expression vector and tested by transfection into the 293FT human cells, with subsequent qPCR analysis of DYRK1A expression.

    Article Title: Hepatitis E Virus Induces Brain Injury Probably Associated With Mitochondrial Apoptosis
    Article Snippet: .. TUNEL Assay TUNEL staining for apoptotic cells in the histological sections was performed using in situ BrdU-Red DNA fragmentation (TUNEL) Assay Kit (ab66110, Abcam) and slides were counterstained with DAPI. ..

    Article Title: Diallyl Sulfide-Mediated Modulation of the Fatty Acid Synthase (FASN) Leads to Cancer Cell Death in BaP-Induced Lung Carcinogenesis in Swiss Mice
    Article Snippet: .. Terminal Transferase-Mediated dUTP Nick End-Labeling (TUNEL) AssayThe apoptosis index in the tissues of treated mice was evaluated using in situ BrdU-Red DNA fragmentation (TUNEL) assay kit from the Abcam following the manufacturer’s instructions. ..

    DNA Fragmentation Assay:

    Article Title: Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments
    Article Snippet: Chemicals Berberine (catalog no: B3251-10G), MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium], propidium iodide (PI), ethidium bromide (EtBr), agarose, proteinase-K protease inhibitor cocktail (catalog no: P8340-5ML) and other accessory chemicals were procured from Sigma (St. Louis, MO). .. An in situ BrdU-Red DNA fragmentation assay kit (TUNEL staining kit, catalog no: ab66110) and Annexin-V staining kit (catalog no: ab176749) were purchased from Abcam (Cambridge, MA). .. Low melting agarose was from Invitrogen (Grand Island, NY; catalog no: 16520050).

    TUNEL Assay:

    Article Title: Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments
    Article Snippet: Chemicals Berberine (catalog no: B3251-10G), MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium], propidium iodide (PI), ethidium bromide (EtBr), agarose, proteinase-K protease inhibitor cocktail (catalog no: P8340-5ML) and other accessory chemicals were procured from Sigma (St. Louis, MO). .. An in situ BrdU-Red DNA fragmentation assay kit (TUNEL staining kit, catalog no: ab66110) and Annexin-V staining kit (catalog no: ab176749) were purchased from Abcam (Cambridge, MA). .. Low melting agarose was from Invitrogen (Grand Island, NY; catalog no: 16520050).

    Article Title: Apoptosis, Induced by Human α-Synuclein in Yeast, Can Occur Independent of Functional Mitochondria
    Article Snippet: .. Ρ− petite cells, which underwent loss of MMP with increasing α-syn copy number, were then assessed via TUNEL assay to determine whether nuclear DNA fragmentation, a hallmark of apoptosis [ ], occurred in these cells. ..

    Article Title: Class I HDAC inhibitor mocetinostat induces apoptosis by activation of miR-31 expression and suppression of E2F6
    Article Snippet: .. For in vivo apoptosis detection, the in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (Abcam, Cambridge, MA, USA) was used following the manufacturer’s protocol. ..

    Article Title: Mutant FUS causes DNA ligation defects to inhibit oxidative damage repair in Amyotrophic Lateral Sclerosis
    Article Snippet: TUNEL staining was used to analyze apoptosis in motor neurons. .. TUNEL Assay Kit-In situ BrdU-Red DNA Fragmentation (Abcam) NeuroTACS II in situ Apoptosis Detection kit (TREVIGEN) was used for the staining. ..

    Article Title: Inhibition of DYRK1A disrupts neural lineage specificationin human pluripotent stem cells
    Article Snippet: The cell cycle distribution of control and ID-8 treated pluripotent WA09 cells was analysed using the Click-IT EdU Flow Cytometry Kit AF488 (Invitrogen/Thermofisher, #C-10425). .. Apoptosis of control and ID-8 treated pluripotent WA09 cells was analysed using the Abcam In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (Abcam, Cambridge, MA). .. To identify gRNAs capable of driving CRISPRa activation of the DYRK1A gene, a number of unique 20mers in region between −250 to −50 bps relative to the main transcription start site were cloned in the pX462 gRNA and dCas9-VP64-expression vector and tested by transfection into the 293FT human cells, with subsequent qPCR analysis of DYRK1A expression.

    Article Title: Hepatitis E Virus Induces Brain Injury Probably Associated With Mitochondrial Apoptosis
    Article Snippet: .. TUNEL Assay TUNEL staining for apoptotic cells in the histological sections was performed using in situ BrdU-Red DNA fragmentation (TUNEL) Assay Kit (ab66110, Abcam) and slides were counterstained with DAPI. ..

    Article Title: Diallyl Sulfide-Mediated Modulation of the Fatty Acid Synthase (FASN) Leads to Cancer Cell Death in BaP-Induced Lung Carcinogenesis in Swiss Mice
    Article Snippet: .. Terminal Transferase-Mediated dUTP Nick End-Labeling (TUNEL) AssayThe apoptosis index in the tissues of treated mice was evaluated using in situ BrdU-Red DNA fragmentation (TUNEL) assay kit from the Abcam following the manufacturer’s instructions. ..

    Article Title: Transthyretin provides trophic support via megalin by promoting neurite outgrowth and neuroprotection in cerebral ischemia
    Article Snippet: .. Primary antibodies used were anti-mouse TTR (1:150, Rabbit, custom made), anti-megalin (1:1000, Sheep, custom made), anti-β III-tubulin (1:500, monoclonal mouse, Promega, Madison, WI, USA, G712A), anti-BrdU (TUNEL Assay Kit, Abcam, ab66110); as secondary antibodies Alexa Fluor 488 and 568 (1:750, Life Technologies) were used. .. The fluorescent dye Hoechst 33342 (0.5 μ g/ml–15 min room temperature) was used to stain nuclei.

    Staining:

    Article Title: Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments
    Article Snippet: Chemicals Berberine (catalog no: B3251-10G), MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium], propidium iodide (PI), ethidium bromide (EtBr), agarose, proteinase-K protease inhibitor cocktail (catalog no: P8340-5ML) and other accessory chemicals were procured from Sigma (St. Louis, MO). .. An in situ BrdU-Red DNA fragmentation assay kit (TUNEL staining kit, catalog no: ab66110) and Annexin-V staining kit (catalog no: ab176749) were purchased from Abcam (Cambridge, MA). .. Low melting agarose was from Invitrogen (Grand Island, NY; catalog no: 16520050).

    Article Title: Mutant FUS causes DNA ligation defects to inhibit oxidative damage repair in Amyotrophic Lateral Sclerosis
    Article Snippet: TUNEL staining was used to analyze apoptosis in motor neurons. .. TUNEL Assay Kit-In situ BrdU-Red DNA Fragmentation (Abcam) NeuroTACS II in situ Apoptosis Detection kit (TREVIGEN) was used for the staining. ..

    Article Title: Hepatitis E Virus Induces Brain Injury Probably Associated With Mitochondrial Apoptosis
    Article Snippet: .. TUNEL Assay TUNEL staining for apoptotic cells in the histological sections was performed using in situ BrdU-Red DNA fragmentation (TUNEL) Assay Kit (ab66110, Abcam) and slides were counterstained with DAPI. ..

    In Vivo:

    Article Title: Class I HDAC inhibitor mocetinostat induces apoptosis by activation of miR-31 expression and suppression of E2F6
    Article Snippet: .. For in vivo apoptosis detection, the in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (Abcam, Cambridge, MA, USA) was used following the manufacturer’s protocol. ..

    End Labeling:

    Article Title: Diallyl Sulfide-Mediated Modulation of the Fatty Acid Synthase (FASN) Leads to Cancer Cell Death in BaP-Induced Lung Carcinogenesis in Swiss Mice
    Article Snippet: .. Terminal Transferase-Mediated dUTP Nick End-Labeling (TUNEL) AssayThe apoptosis index in the tissues of treated mice was evaluated using in situ BrdU-Red DNA fragmentation (TUNEL) assay kit from the Abcam following the manufacturer’s instructions. ..

    Mouse Assay:

    Article Title: Diallyl Sulfide-Mediated Modulation of the Fatty Acid Synthase (FASN) Leads to Cancer Cell Death in BaP-Induced Lung Carcinogenesis in Swiss Mice
    Article Snippet: .. Terminal Transferase-Mediated dUTP Nick End-Labeling (TUNEL) AssayThe apoptosis index in the tissues of treated mice was evaluated using in situ BrdU-Red DNA fragmentation (TUNEL) assay kit from the Abcam following the manufacturer’s instructions. ..

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    Abcam in situ brdu red dna fragmentation tunel assay kit
    More <t>DNA</t> fragmention of U-87MG and U-251 cells occurred after the combined SLCP and BBR therapy than by individual treatments. U-87MG and U-251MG cells were grown in EMEM and they were treated with either SLCP (20 μM), BBR (100 μM) or their combination (in the 1:5 ratio) for 48 h. The cells were stained with terminal deoxyribonucleic acid nick end labeling <t>(TUNEL)</t> reagents for detecting DNA fragmented cells and counter-stained with Hoechst 33342. A fluorescent microscope (Leica Germany) was used to detect the signal with appropriate excitation/emission filters. A-B: Representative images of TUNEL/Hoechst 33342 stained U-87MG (A) and U-251MG (B) cells after treatment with SLCP, BBR or their combination. C-D: More TUNEL-positive cells were observed in combination-treated cells in both GB cell lines than for individual treatments. Arrows indicate TUNEL positive cells. Scale bars indicate 100 μm and is applicable to all images. *p
    In Situ Brdu Red Dna Fragmentation Tunel Assay Kit, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/in situ brdu red dna fragmentation tunel assay kit/product/Abcam
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    in situ brdu red dna fragmentation tunel assay kit - by Bioz Stars, 2021-03
    95/100 stars
      Buy from Supplier

    Image Search Results


    More DNA fragmention of U-87MG and U-251 cells occurred after the combined SLCP and BBR therapy than by individual treatments. U-87MG and U-251MG cells were grown in EMEM and they were treated with either SLCP (20 μM), BBR (100 μM) or their combination (in the 1:5 ratio) for 48 h. The cells were stained with terminal deoxyribonucleic acid nick end labeling (TUNEL) reagents for detecting DNA fragmented cells and counter-stained with Hoechst 33342. A fluorescent microscope (Leica Germany) was used to detect the signal with appropriate excitation/emission filters. A-B: Representative images of TUNEL/Hoechst 33342 stained U-87MG (A) and U-251MG (B) cells after treatment with SLCP, BBR or their combination. C-D: More TUNEL-positive cells were observed in combination-treated cells in both GB cell lines than for individual treatments. Arrows indicate TUNEL positive cells. Scale bars indicate 100 μm and is applicable to all images. *p

    Journal: PLoS ONE

    Article Title: Combination treatment of berberine and solid lipid curcumin particles increased cell death and inhibited PI3K/Akt/mTOR pathway of human cultured glioblastoma cells more effectively than did individual treatments

    doi: 10.1371/journal.pone.0225660

    Figure Lengend Snippet: More DNA fragmention of U-87MG and U-251 cells occurred after the combined SLCP and BBR therapy than by individual treatments. U-87MG and U-251MG cells were grown in EMEM and they were treated with either SLCP (20 μM), BBR (100 μM) or their combination (in the 1:5 ratio) for 48 h. The cells were stained with terminal deoxyribonucleic acid nick end labeling (TUNEL) reagents for detecting DNA fragmented cells and counter-stained with Hoechst 33342. A fluorescent microscope (Leica Germany) was used to detect the signal with appropriate excitation/emission filters. A-B: Representative images of TUNEL/Hoechst 33342 stained U-87MG (A) and U-251MG (B) cells after treatment with SLCP, BBR or their combination. C-D: More TUNEL-positive cells were observed in combination-treated cells in both GB cell lines than for individual treatments. Arrows indicate TUNEL positive cells. Scale bars indicate 100 μm and is applicable to all images. *p

    Article Snippet: An in situ BrdU-Red DNA fragmentation assay kit (TUNEL staining kit, catalog no: ab66110) and Annexin-V staining kit (catalog no: ab176749) were purchased from Abcam (Cambridge, MA).

    Techniques: Staining, End Labeling, TUNEL Assay, Microscopy

    ( A ) Representative pictures of growth of ρ 0 yeast cells harboring 1-copy (1 C ), 2-copies (2 C ), and 3-copies (3 C ) of the human α-syn gene, under the control of either MET25 p, on complete YPD medium agar plates in the presence (YPD + Methionine) or absence (YPD) of methionine. ( B ) Western blot analyses of ρ 0 cells expressing 1-copy (lane 1), 2-copies (lane 2), and 3-copies (lane 3) of α-syn protein. On lanes 1, 2, and 3 were loaded 7.5 µg of total protein obtained after lysis of cells. The upper panel was probed with an antibody that recognizes the HA epitope (Proteintech, 51064-2-AP) and the lower panel with a β-actin antibody (Proteintech, 60008-1-Ig); levels of β-actin were used as loading controls, β-actin being a housekeeping gene. ( C ) Microscopic images (×400) of ρ 0 cells, stained with Phloxine B, after the expression of 1–3 copies of α-syn from the MET25 p. ( D ) Microscopic images (×400) of nuclear DNA fragmentation, as observed using the TUNEL assay, in ρ 0 yeast cells expressing 1–3-copies of α-syn from MET25 p. ( E ) ρ 0 cells, bearing 1–3 copies of MET25 p-driven α-syn expression cassettes, were stained with Hoechst 33,342 (a blue dye that labels DNA of live cells). Post Hoc Newman-Keuls test after a significant one-way ANOVA test, indicated a significant difference between petites expressing 1–3 copies of α-syn.

    Journal: Cells

    Article Title: Apoptosis, Induced by Human α-Synuclein in Yeast, Can Occur Independent of Functional Mitochondria

    doi: 10.3390/cells9102203

    Figure Lengend Snippet: ( A ) Representative pictures of growth of ρ 0 yeast cells harboring 1-copy (1 C ), 2-copies (2 C ), and 3-copies (3 C ) of the human α-syn gene, under the control of either MET25 p, on complete YPD medium agar plates in the presence (YPD + Methionine) or absence (YPD) of methionine. ( B ) Western blot analyses of ρ 0 cells expressing 1-copy (lane 1), 2-copies (lane 2), and 3-copies (lane 3) of α-syn protein. On lanes 1, 2, and 3 were loaded 7.5 µg of total protein obtained after lysis of cells. The upper panel was probed with an antibody that recognizes the HA epitope (Proteintech, 51064-2-AP) and the lower panel with a β-actin antibody (Proteintech, 60008-1-Ig); levels of β-actin were used as loading controls, β-actin being a housekeeping gene. ( C ) Microscopic images (×400) of ρ 0 cells, stained with Phloxine B, after the expression of 1–3 copies of α-syn from the MET25 p. ( D ) Microscopic images (×400) of nuclear DNA fragmentation, as observed using the TUNEL assay, in ρ 0 yeast cells expressing 1–3-copies of α-syn from MET25 p. ( E ) ρ 0 cells, bearing 1–3 copies of MET25 p-driven α-syn expression cassettes, were stained with Hoechst 33,342 (a blue dye that labels DNA of live cells). Post Hoc Newman-Keuls test after a significant one-way ANOVA test, indicated a significant difference between petites expressing 1–3 copies of α-syn.

    Article Snippet: Ρ− petite cells, which underwent loss of MMP with increasing α-syn copy number, were then assessed via TUNEL assay to determine whether nuclear DNA fragmentation, a hallmark of apoptosis [ ], occurred in these cells.

    Techniques: Western Blot, Expressing, Lysis, Staining, TUNEL Assay

    Analysis of nuclear DNA fragmentation ( A ), an increase of reactive oxygen species (ROS; B ), and loss of mitochondrial membrane potential (MMP; C ), in ρ 0 yeast cells that bear 1 to 3-copies of the human α-syn, gene under the control of the MET25 promoter. ( A ) Levels of nuclear DNA fragmentation, a hallmark of apoptosis, in yeast cells that express 1 to 3-copies of α-syn were assessed using the TUNEL assay. ( B ) Cells were stained with dihydroethidine to detect ROS. ( C ) Cells were stained with the JC-10 dye to quantify MMP loss. The data presented in ( A , B , C ) represent mean ± S.D. of three independent experiments ( p

    Journal: Cells

    Article Title: Apoptosis, Induced by Human α-Synuclein in Yeast, Can Occur Independent of Functional Mitochondria

    doi: 10.3390/cells9102203

    Figure Lengend Snippet: Analysis of nuclear DNA fragmentation ( A ), an increase of reactive oxygen species (ROS; B ), and loss of mitochondrial membrane potential (MMP; C ), in ρ 0 yeast cells that bear 1 to 3-copies of the human α-syn, gene under the control of the MET25 promoter. ( A ) Levels of nuclear DNA fragmentation, a hallmark of apoptosis, in yeast cells that express 1 to 3-copies of α-syn were assessed using the TUNEL assay. ( B ) Cells were stained with dihydroethidine to detect ROS. ( C ) Cells were stained with the JC-10 dye to quantify MMP loss. The data presented in ( A , B , C ) represent mean ± S.D. of three independent experiments ( p

    Article Snippet: Ρ− petite cells, which underwent loss of MMP with increasing α-syn copy number, were then assessed via TUNEL assay to determine whether nuclear DNA fragmentation, a hallmark of apoptosis [ ], occurred in these cells.

    Techniques: TUNEL Assay, Staining

    TTR neuroprotection in vivo (pMCAO) is also megalin-dependent. ( a ) Megalin levels after pMCAO were accessed by immunofluorescence confocal stacks of the different brain areas of wild-type and TTR KO mice (HSF+/− background) after 24 h of pMCAO. Representative immunofluorescence confocal stack of megalin in infarct area (IF Ipsi) and contralateral area (IF Contra) (scale bar 50 μ m). ( b ) The semi-quantification results are the average±S.E.M. of three to five stacks/animal (three animals for each phenotype). ( c ) Megalin mRNA levels were quantified for the different brain areas (IF Contra versus IF Ipsi; P-IF Contra versus P-IF Ipsi) of the 24 h pMCAO WT and TTR KO mice (HSF+/− background). The results are the average±S.E.M. of three to four different animals for each phenotype. ( d ) Megalin protein levels were quantified for the different brain areas (IF Contra versus IF Ipsi) of the 24 h pMCAO WT and TTR KO mice (HSF+/− background). The results are the average±S.E.M. of three different animals for each phenotype. ( e ) Representative images of megalin immunofluorescence confocal stacks of live cells co-localizing with β III-tubulin from the infarct area of WT mice (HSF+/− background) after 24 h pMCAO (three animals). ( f and g ) Semi-quantification of the cell survival accessed with TUNEL reaction assay from the megalin-positive cells in the infarct area of TTR KO and WT mice (HSF+/− background) after 24 h pMCAO. The results are the average±S.E.M. of five to six stacks from three animals for each phenotype ( f , representative image; g ). ( h ) Representative immunofluorescence confocal stack of TTR-positive live cells co-localizing with megalin, from the infarct area of WT mice (HSF+/− background), after 24 h pMCAO (three animals). ( i and j ) Semi-quantification of the cell survival from the TTR-positive cells in the infarct area of WT mice (HSF+/− background) after 24 h pMCAO. Cell survival was accessed with TUNEL reaction assay. The results are the average±S.E.M. of six stacks from three WT pMCAO animals ( i , representative images; j ). Statistical analysis was performed using one-way ANOVA followed by Bonferroni's multiple comparison test performed for each condition or Student's unpaired t -test for two-group comparisons. *** P

    Journal: Cell Death and Differentiation

    Article Title: Transthyretin provides trophic support via megalin by promoting neurite outgrowth and neuroprotection in cerebral ischemia

    doi: 10.1038/cdd.2016.64

    Figure Lengend Snippet: TTR neuroprotection in vivo (pMCAO) is also megalin-dependent. ( a ) Megalin levels after pMCAO were accessed by immunofluorescence confocal stacks of the different brain areas of wild-type and TTR KO mice (HSF+/− background) after 24 h of pMCAO. Representative immunofluorescence confocal stack of megalin in infarct area (IF Ipsi) and contralateral area (IF Contra) (scale bar 50 μ m). ( b ) The semi-quantification results are the average±S.E.M. of three to five stacks/animal (three animals for each phenotype). ( c ) Megalin mRNA levels were quantified for the different brain areas (IF Contra versus IF Ipsi; P-IF Contra versus P-IF Ipsi) of the 24 h pMCAO WT and TTR KO mice (HSF+/− background). The results are the average±S.E.M. of three to four different animals for each phenotype. ( d ) Megalin protein levels were quantified for the different brain areas (IF Contra versus IF Ipsi) of the 24 h pMCAO WT and TTR KO mice (HSF+/− background). The results are the average±S.E.M. of three different animals for each phenotype. ( e ) Representative images of megalin immunofluorescence confocal stacks of live cells co-localizing with β III-tubulin from the infarct area of WT mice (HSF+/− background) after 24 h pMCAO (three animals). ( f and g ) Semi-quantification of the cell survival accessed with TUNEL reaction assay from the megalin-positive cells in the infarct area of TTR KO and WT mice (HSF+/− background) after 24 h pMCAO. The results are the average±S.E.M. of five to six stacks from three animals for each phenotype ( f , representative image; g ). ( h ) Representative immunofluorescence confocal stack of TTR-positive live cells co-localizing with megalin, from the infarct area of WT mice (HSF+/− background), after 24 h pMCAO (three animals). ( i and j ) Semi-quantification of the cell survival from the TTR-positive cells in the infarct area of WT mice (HSF+/− background) after 24 h pMCAO. Cell survival was accessed with TUNEL reaction assay. The results are the average±S.E.M. of six stacks from three WT pMCAO animals ( i , representative images; j ). Statistical analysis was performed using one-way ANOVA followed by Bonferroni's multiple comparison test performed for each condition or Student's unpaired t -test for two-group comparisons. *** P

    Article Snippet: Primary antibodies used were anti-mouse TTR (1:150, Rabbit, custom made), anti-megalin (1:1000, Sheep, custom made), anti-β III-tubulin (1:500, monoclonal mouse, Promega, Madison, WI, USA, G712A), anti-BrdU (TUNEL Assay Kit, Abcam, ab66110); as secondary antibodies Alexa Fluor 488 and 568 (1:750, Life Technologies) were used.

    Techniques: In Vivo, Immunofluorescence, Mouse Assay, TUNEL Assay

    TUNEL-positive cells increased after HEV infection. TUNEL staining of the HEV-RNA positive brain tissues collected on 14, 21, and 28 dpi was performed using in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit and slides were counterstained with DAPI (Blue). TUNEL-positive signals merged with DAPI were shown as purple. (A,B) Few positive signals were detected in animals of mock group. (C–E) Positive signals were exhibited in infected brain sections. The high magnification images (D,E) depicted granular masses of positive signals at nucleus of the apoptotic cells. (F) TUNEL-positive cells were significantly increased in HEV-infected gerbil brain tissues (* p

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Hepatitis E Virus Induces Brain Injury Probably Associated With Mitochondrial Apoptosis

    doi: 10.3389/fcimb.2019.00433

    Figure Lengend Snippet: TUNEL-positive cells increased after HEV infection. TUNEL staining of the HEV-RNA positive brain tissues collected on 14, 21, and 28 dpi was performed using in situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit and slides were counterstained with DAPI (Blue). TUNEL-positive signals merged with DAPI were shown as purple. (A,B) Few positive signals were detected in animals of mock group. (C–E) Positive signals were exhibited in infected brain sections. The high magnification images (D,E) depicted granular masses of positive signals at nucleus of the apoptotic cells. (F) TUNEL-positive cells were significantly increased in HEV-infected gerbil brain tissues (* p

    Article Snippet: TUNEL Assay TUNEL staining for apoptotic cells in the histological sections was performed using in situ BrdU-Red DNA fragmentation (TUNEL) Assay Kit (ab66110, Abcam) and slides were counterstained with DAPI.

    Techniques: TUNEL Assay, Infection, Staining, In Situ