Structured Review

Trevigen dna fragmentation
Immunoelectron microscopic characterization of apoptosis at the rupture sites. A: Ultrastructural examination of the macrophages ( arrows ) at the rupture site showed that the majority had nuclear changes suggestive of apoptosis. B: Example demonstrating varying stages of nuclear shrinkage with condensation of chromatin at the nuclear periphery. The presence of lipid globules and lack of contractile elements help identify the macrophages. ISEL was performed on ultra-thin sections with <t>TdT</t> and biotinylated nucleotides. Fragmented <t>DNA</t> was visualized using a 10-nm gold, streptavidin conjugate. C: High-power electron micrograph of the region indicated by the inset in B , note the gold particles are selectively localized on the condensed chromatin material.
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Images

1) Product Images from "Localization of Apoptotic Macrophages at the Site of Plaque Rupture in Sudden Coronary Death"

Article Title: Localization of Apoptotic Macrophages at the Site of Plaque Rupture in Sudden Coronary Death

Journal: The American Journal of Pathology

doi:

Immunoelectron microscopic characterization of apoptosis at the rupture sites. A: Ultrastructural examination of the macrophages ( arrows ) at the rupture site showed that the majority had nuclear changes suggestive of apoptosis. B: Example demonstrating varying stages of nuclear shrinkage with condensation of chromatin at the nuclear periphery. The presence of lipid globules and lack of contractile elements help identify the macrophages. ISEL was performed on ultra-thin sections with TdT and biotinylated nucleotides. Fragmented DNA was visualized using a 10-nm gold, streptavidin conjugate. C: High-power electron micrograph of the region indicated by the inset in B , note the gold particles are selectively localized on the condensed chromatin material.
Figure Legend Snippet: Immunoelectron microscopic characterization of apoptosis at the rupture sites. A: Ultrastructural examination of the macrophages ( arrows ) at the rupture site showed that the majority had nuclear changes suggestive of apoptosis. B: Example demonstrating varying stages of nuclear shrinkage with condensation of chromatin at the nuclear periphery. The presence of lipid globules and lack of contractile elements help identify the macrophages. ISEL was performed on ultra-thin sections with TdT and biotinylated nucleotides. Fragmented DNA was visualized using a 10-nm gold, streptavidin conjugate. C: High-power electron micrograph of the region indicated by the inset in B , note the gold particles are selectively localized on the condensed chromatin material.

Techniques Used:

2) Product Images from "Intracellular Amyloidogenesis by Human Islet Amyloid Polypeptide Induces Apoptosis in COS-1 Cells"

Article Title: Intracellular Amyloidogenesis by Human Islet Amyloid Polypeptide Induces Apoptosis in COS-1 Cells

Journal: The American Journal of Pathology

doi:

Double labeling of intracellular hIAPP and specific markers of apoptosis is found only in hIAPP-expressing COS-1 cells. A: Intracellular accumulation of hIAPP is associated with extracellular annexin-V labeling indicative of apoptosis in pMT2-hIAPP -transfected COS-1 cells. Extracellular annexin was fluorescently labeled with streptavidin-rhodamine ( red ) and intracellular IAPP with FITC ( green ). The majority ( > 90%) of control pMT2-hIAPP anti -transfected cells ( B ) have only background levels of annexin labeling; the single brightly labeled cell in B is undergoing apoptosis. Intracellular accumulation of hIAPP ( green ) is also associated with TUNEL labeling of fragmented DNA ( red ) in the nucleus of pMT2-hIAPP - ( C ) but not control pMT2 -transfected cells ( D ). Double labeling of adherent COS-1 cells was performed as described in Materials and Methods.
Figure Legend Snippet: Double labeling of intracellular hIAPP and specific markers of apoptosis is found only in hIAPP-expressing COS-1 cells. A: Intracellular accumulation of hIAPP is associated with extracellular annexin-V labeling indicative of apoptosis in pMT2-hIAPP -transfected COS-1 cells. Extracellular annexin was fluorescently labeled with streptavidin-rhodamine ( red ) and intracellular IAPP with FITC ( green ). The majority ( > 90%) of control pMT2-hIAPP anti -transfected cells ( B ) have only background levels of annexin labeling; the single brightly labeled cell in B is undergoing apoptosis. Intracellular accumulation of hIAPP ( green ) is also associated with TUNEL labeling of fragmented DNA ( red ) in the nucleus of pMT2-hIAPP - ( C ) but not control pMT2 -transfected cells ( D ). Double labeling of adherent COS-1 cells was performed as described in Materials and Methods.

Techniques Used: Labeling, Expressing, Transfection, TUNEL Assay

3) Product Images from "A Study of the Interferon Antiviral Mechanism: Apoptosis Activation by the 2-5A System "

Article Title: A Study of the Interferon Antiviral Mechanism: Apoptosis Activation by the 2-5A System

Journal: The Journal of Experimental Medicine

doi:

RNase L overexpression caused DNA cleavage characteristic of apoptosis. 3T3/neo ( A and C ) and 3T3/RNaseLS ( B and D ) cells were incubated for 24 h in the absence ( A and B ) and presence ( C and D ) of 3 mM IPTG. Apoptotic cells were detected in situ by using T7 DNA polymerase with methyl green counterstaining. Experiments were performed in triplicate.
Figure Legend Snippet: RNase L overexpression caused DNA cleavage characteristic of apoptosis. 3T3/neo ( A and C ) and 3T3/RNaseLS ( B and D ) cells were incubated for 24 h in the absence ( A and B ) and presence ( C and D ) of 3 mM IPTG. Apoptotic cells were detected in situ by using T7 DNA polymerase with methyl green counterstaining. Experiments were performed in triplicate.

Techniques Used: Over Expression, Incubation, In Situ

4) Product Images from "Salinomycin inhibits metastatic colorectal cancer growth and interferes with Wnt/β-catenin signaling in CD133+ human colorectal cancer cells"

Article Title: Salinomycin inhibits metastatic colorectal cancer growth and interferes with Wnt/β-catenin signaling in CD133+ human colorectal cancer cells

Journal: BMC Cancer

doi: 10.1186/s12885-016-2879-8

Induction of cell death of murine colorectal cancer cells after treatment with Salinomycin. 0.5 × 10 6 MC38 ( a ) or CT26 ( b ) cells were seeded in 6-well plates and grown until confluence following exposure to 1 μM 5-FU and increasing concentrations of Salinomycin (1 μM, 2 μM, 5 μM and 10 μM) for 24 h. Detection of cell death was performed using AnnexinV-FITC and PI staining and cells analyzed by flowcytometry. Cell death was further determined by quantification of DNA fragmentation ( c + d ), c-PARP production ( e + f ) and LDH release assay ( g + h ). Results are displayed as representative dot blots or as summary of at least 3 independent experiments; * p
Figure Legend Snippet: Induction of cell death of murine colorectal cancer cells after treatment with Salinomycin. 0.5 × 10 6 MC38 ( a ) or CT26 ( b ) cells were seeded in 6-well plates and grown until confluence following exposure to 1 μM 5-FU and increasing concentrations of Salinomycin (1 μM, 2 μM, 5 μM and 10 μM) for 24 h. Detection of cell death was performed using AnnexinV-FITC and PI staining and cells analyzed by flowcytometry. Cell death was further determined by quantification of DNA fragmentation ( c + d ), c-PARP production ( e + f ) and LDH release assay ( g + h ). Results are displayed as representative dot blots or as summary of at least 3 independent experiments; * p

Techniques Used: Staining, Lactate Dehydrogenase Assay

5) Product Images from "Hemin Causes Lung Microvascular Endothelial Barrier Dysfunction by Necroptotic Cell Death"

Article Title: Hemin Causes Lung Microvascular Endothelial Barrier Dysfunction by Necroptotic Cell Death

Journal: American Journal of Respiratory Cell and Molecular Biology

doi: 10.1165/rcmb.2016-0287OC

Treatment with 40 μM hemin causes programmed hLMVEC death. hLMVECs grown to confluence in 6-well plates were treated with either vehicle or 40 μM hemin. Cells were trypsinized and stained with Trypan blue. Aliquots of the stained cell suspensions were placed in a hemocytometer chamber on a slide and visualized under 20× magnification with a light microscope. Representative fields for each condition are shown in A . In B , the bar graph depicts the average percentage of Trypan-blue–positive cells in 10 high-powered fields for control versus hemin-treated groups. In separate experiments, hLMVECs were grown in 96-well plates and treated with 400 nM TAK-242 and/or 1 mM NAC or 1 μM deferoxamine versus vehicle controls + 40 μM hemin. DNA fragmentation was detected in situ by TdT nick-end labeling with biotinylated nucleotides, followed by secondary labeling with streptavidin-horseradish peroxidase and incubation with peroxidase substrates, resulting in a colorimetric readout quantified by spectrophotometric absorbance at 450 nm. The bar graph in C depicts the average relative absorbances of the various treatment conditions. Error bars are ± SD. *Indicates significant difference from control (Student’s t test, P
Figure Legend Snippet: Treatment with 40 μM hemin causes programmed hLMVEC death. hLMVECs grown to confluence in 6-well plates were treated with either vehicle or 40 μM hemin. Cells were trypsinized and stained with Trypan blue. Aliquots of the stained cell suspensions were placed in a hemocytometer chamber on a slide and visualized under 20× magnification with a light microscope. Representative fields for each condition are shown in A . In B , the bar graph depicts the average percentage of Trypan-blue–positive cells in 10 high-powered fields for control versus hemin-treated groups. In separate experiments, hLMVECs were grown in 96-well plates and treated with 400 nM TAK-242 and/or 1 mM NAC or 1 μM deferoxamine versus vehicle controls + 40 μM hemin. DNA fragmentation was detected in situ by TdT nick-end labeling with biotinylated nucleotides, followed by secondary labeling with streptavidin-horseradish peroxidase and incubation with peroxidase substrates, resulting in a colorimetric readout quantified by spectrophotometric absorbance at 450 nm. The bar graph in C depicts the average relative absorbances of the various treatment conditions. Error bars are ± SD. *Indicates significant difference from control (Student’s t test, P

Techniques Used: Staining, Light Microscopy, In Situ, End Labeling, Labeling, Incubation

6) Product Images from "Phosphorylation of CLK2 at Serine 34 and Threonine 127 by AKT Controls Cell Survival after Ionizing Radiation *"

Article Title: Phosphorylation of CLK2 at Serine 34 and Threonine 127 by AKT Controls Cell Survival after Ionizing Radiation *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M110.122044

The effects of CLK2 on radiation sensitivity of CCD-18Lu cells. Cells were transfected with either vacant vector or myc-CLK2 . At 24 h post-transfection, cells were exposed to 0.05–2 Gy of γ-rays and cell viability was assayed 48 h later. A , the overexpression of Myc-CLK2 at 72 h post-transfection. B , the cell viability of CLK2 overexpressing CCD-18Lu cells to ionizing radiation was determined by MTT assay. C , cell proliferation of CLK2 overexpressing cells after irradiation. D , effect of CLK2 overexpression on 2 Gy-induced apoptosis. DNA fragmentation was detected with HT TiterTACS Assay Kit for the quantification of apoptosis. B–D , data represent mean ± S.D. ( n = 3) and were analyzed by the t test. Data showed a significant difference compared with vector control at the indicated dose (*, p
Figure Legend Snippet: The effects of CLK2 on radiation sensitivity of CCD-18Lu cells. Cells were transfected with either vacant vector or myc-CLK2 . At 24 h post-transfection, cells were exposed to 0.05–2 Gy of γ-rays and cell viability was assayed 48 h later. A , the overexpression of Myc-CLK2 at 72 h post-transfection. B , the cell viability of CLK2 overexpressing CCD-18Lu cells to ionizing radiation was determined by MTT assay. C , cell proliferation of CLK2 overexpressing cells after irradiation. D , effect of CLK2 overexpression on 2 Gy-induced apoptosis. DNA fragmentation was detected with HT TiterTACS Assay Kit for the quantification of apoptosis. B–D , data represent mean ± S.D. ( n = 3) and were analyzed by the t test. Data showed a significant difference compared with vector control at the indicated dose (*, p

Techniques Used: Transfection, Plasmid Preparation, Over Expression, MTT Assay, Irradiation

The regulation of the sensitivity of CCD-18Lu cells to ionizing radiation is dependent on CLK2 phosphorylation. A , CCD-18Lu cells were transfected with vacant vector, WT- CLK2 , or CLK2 mutants and the overexpressions of WT-CLK2 and CLK2 mutants were detected. B , the viabilities of vector, WT-CLK2, or mutant-CLK2 overexpressing cells were measured using MTT assays 48 h after 2 Gy of γ-ray. C , effect of CLK2 phosphorylation on 2 Gy-induced apoptosis. DNA fragmentation was detected with HT TiterTACS Assay Kit for the quantification of apoptosis. The absorbance (450 nm) of untreated vector control was 0.34. D , the viabilities of vacant vector, WT-CLK2, or mutant-CLK2 overexpressing cells after exposure to 0.05 Gy of γ-ray were measured by MTT assay. E , the cell proliferation after irradiation of 0.05 Gy in vacant vector, WT-CLK2, or mutant-CLK2 CCD-18Lu cells. Data represent mean ± S.D. ( n = 3) and were analyzed by Dunnett's test for multiple comparison in analysis of variance to irradiated vector control (*, p
Figure Legend Snippet: The regulation of the sensitivity of CCD-18Lu cells to ionizing radiation is dependent on CLK2 phosphorylation. A , CCD-18Lu cells were transfected with vacant vector, WT- CLK2 , or CLK2 mutants and the overexpressions of WT-CLK2 and CLK2 mutants were detected. B , the viabilities of vector, WT-CLK2, or mutant-CLK2 overexpressing cells were measured using MTT assays 48 h after 2 Gy of γ-ray. C , effect of CLK2 phosphorylation on 2 Gy-induced apoptosis. DNA fragmentation was detected with HT TiterTACS Assay Kit for the quantification of apoptosis. The absorbance (450 nm) of untreated vector control was 0.34. D , the viabilities of vacant vector, WT-CLK2, or mutant-CLK2 overexpressing cells after exposure to 0.05 Gy of γ-ray were measured by MTT assay. E , the cell proliferation after irradiation of 0.05 Gy in vacant vector, WT-CLK2, or mutant-CLK2 CCD-18Lu cells. Data represent mean ± S.D. ( n = 3) and were analyzed by Dunnett's test for multiple comparison in analysis of variance to irradiated vector control (*, p

Techniques Used: Transfection, Plasmid Preparation, Mutagenesis, MTT Assay, Irradiation

7) Product Images from "Genotoxicants Target Distinct Molecular Networks in Neonatal Neurons"

Article Title: Genotoxicants Target Distinct Molecular Networks in Neonatal Neurons

Journal: Environmental Health Perspectives

doi: 10.1289/ehp.9073

In situ DNA damage of cerebellar neurons treated with MAM or HN2. ( A–C ) Representative light micrographs of cerebellar neurons treated for 24 hr with various concentrations of MAM or HN2 and examined for the extent of DNA fragmentation by TUNEL labeling ( A ) or N 7-alkylguanine DNA lesions induced by 100 μM MAM ( B ) or 0.1–10 μM HN2 ( C ). Note the extensive labeling of neurons treated with 10μM HN2 or 1,000 μM MAM. Bar = 50 μm. Significantly different from control-treated neurons (* p
Figure Legend Snippet: In situ DNA damage of cerebellar neurons treated with MAM or HN2. ( A–C ) Representative light micrographs of cerebellar neurons treated for 24 hr with various concentrations of MAM or HN2 and examined for the extent of DNA fragmentation by TUNEL labeling ( A ) or N 7-alkylguanine DNA lesions induced by 100 μM MAM ( B ) or 0.1–10 μM HN2 ( C ). Note the extensive labeling of neurons treated with 10μM HN2 or 1,000 μM MAM. Bar = 50 μm. Significantly different from control-treated neurons (* p

Techniques Used: In Situ, TUNEL Assay, Labeling

8) Product Images from "Salinomycin inhibits cholangiocarcinoma growth by inhibition of autophagic flux"

Article Title: Salinomycin inhibits cholangiocarcinoma growth by inhibition of autophagic flux

Journal: Oncotarget

doi: 10.18632/oncotarget.23339

Treatment with Salinomycin induces apoptosis in murine CC cells ( A ) A total of 0.5 × 10 6 p246 or p254 cells were seeded in six-well plates and grown until confluence following exposure to increasing concentrations of Salinomycin (1, 2, 5, and 10 µM) for 24 h. Detection of apoptosis was performed using AnnexinV-FITC and propidium iodide staining, and cells were analyzed by flow cytometry. Cell death was further determined by quantification of DNA fragmentation ( B ) and LDH release assay ( C ). Results are displayed as representative dot blots or as a summary of at least three independent experiments; * P
Figure Legend Snippet: Treatment with Salinomycin induces apoptosis in murine CC cells ( A ) A total of 0.5 × 10 6 p246 or p254 cells were seeded in six-well plates and grown until confluence following exposure to increasing concentrations of Salinomycin (1, 2, 5, and 10 µM) for 24 h. Detection of apoptosis was performed using AnnexinV-FITC and propidium iodide staining, and cells were analyzed by flow cytometry. Cell death was further determined by quantification of DNA fragmentation ( B ) and LDH release assay ( C ). Results are displayed as representative dot blots or as a summary of at least three independent experiments; * P

Techniques Used: Staining, Flow Cytometry, Cytometry, Lactate Dehydrogenase Assay

9) Product Images from "Cardioprotective Effect of Tangeretin by Inhibiting PTEN/AKT/mTOR Axis in Experimental Sepsis-Induced Myocardial Dysfunction"

Article Title: Cardioprotective Effect of Tangeretin by Inhibiting PTEN/AKT/mTOR Axis in Experimental Sepsis-Induced Myocardial Dysfunction

Journal: Molecules

doi: 10.3390/molecules25235622

( a ) Representative fluorescent images of TUNEL staining performed on paraffin section of rat heart in each group; Scale bar 5 mm= 25 µm. Green color demonstrated TUNEL-positive nuclei and intensity of the same is plotted as bar graph near the TUNEL images. ( b ) Cardiac cell death markers DNA fragmentation and ( c ) poly(ADP-ribose) polymerase (PARP) activity assay. Both DNA fragmentation and PARP activity were significantly increased in CLP-rats, and TG (50 and 100 mg/kg) treatment ameliorated elevated levels, dose-dependently. Values represented as means ± SD and n = 12/group. One-way analysis of variance (ANOVA), followed by Tukey’s test were used for statically analysis (Significance level * p
Figure Legend Snippet: ( a ) Representative fluorescent images of TUNEL staining performed on paraffin section of rat heart in each group; Scale bar 5 mm= 25 µm. Green color demonstrated TUNEL-positive nuclei and intensity of the same is plotted as bar graph near the TUNEL images. ( b ) Cardiac cell death markers DNA fragmentation and ( c ) poly(ADP-ribose) polymerase (PARP) activity assay. Both DNA fragmentation and PARP activity were significantly increased in CLP-rats, and TG (50 and 100 mg/kg) treatment ameliorated elevated levels, dose-dependently. Values represented as means ± SD and n = 12/group. One-way analysis of variance (ANOVA), followed by Tukey’s test were used for statically analysis (Significance level * p

Techniques Used: TUNEL Assay, Staining, Paraffin Section, Activity Assay

10) Product Images from "Proinflammatory consequences of transgenic Fas ligand expression in the heart"

Article Title: Proinflammatory consequences of transgenic Fas ligand expression in the heart

Journal: Journal of Clinical Investigation

doi:

Cardiomyocyte apoptosis was not detected in 12-week-old FasL Tg hearts by TUNEL or DNA ladder analysis. Paraffin sections of NTg ( a ) and line 61 FasL Tg ( b ) ventricle were stained for apoptotic cells through the labeling of nuclear DNA fragments. The inset in a displays DNase-treated NTg myocardium as a positive control for the TUNEL stain. TUNEL analysis demonstrates no apoptotic or necrotic cardiomyocytes in either NTg or Tg myocardium. Small numbers of TUNEL-positive leukocyte nuclei (arrows) are seen in the interstitium of Tg hearts. The scattered leukocytes, predominantly neutrophils and lymphocytes, are presumably apoptotic. DNA laddering was not detected in NTg, line 70, or line 61 FasL Tg hearts ( c ). Staurosporine-treated rat cardiomyocytes served as positive control for DNA ladder analysis.
Figure Legend Snippet: Cardiomyocyte apoptosis was not detected in 12-week-old FasL Tg hearts by TUNEL or DNA ladder analysis. Paraffin sections of NTg ( a ) and line 61 FasL Tg ( b ) ventricle were stained for apoptotic cells through the labeling of nuclear DNA fragments. The inset in a displays DNase-treated NTg myocardium as a positive control for the TUNEL stain. TUNEL analysis demonstrates no apoptotic or necrotic cardiomyocytes in either NTg or Tg myocardium. Small numbers of TUNEL-positive leukocyte nuclei (arrows) are seen in the interstitium of Tg hearts. The scattered leukocytes, predominantly neutrophils and lymphocytes, are presumably apoptotic. DNA laddering was not detected in NTg, line 70, or line 61 FasL Tg hearts ( c ). Staurosporine-treated rat cardiomyocytes served as positive control for DNA ladder analysis.

Techniques Used: TUNEL Assay, Staining, Labeling, Positive Control, DNA Laddering

11) Product Images from "Induction of apoptosis by pinostrobin in human cervical cancer cells: Possible mechanism of action"

Article Title: Induction of apoptosis by pinostrobin in human cervical cancer cells: Possible mechanism of action

Journal: PLoS ONE

doi: 10.1371/journal.pone.0191523

Cell cycle analysis of apoptotic HeLa cells was performed by flow cytometry after propidium iodide staining. G1/S phase of cell cycle was found arrested by pinostrobin treatment at 48 h. The X-axis (FL-2H) represents DNA content and Y-axis (counts) shows the number of cells in each phase of cell cycle.
Figure Legend Snippet: Cell cycle analysis of apoptotic HeLa cells was performed by flow cytometry after propidium iodide staining. G1/S phase of cell cycle was found arrested by pinostrobin treatment at 48 h. The X-axis (FL-2H) represents DNA content and Y-axis (counts) shows the number of cells in each phase of cell cycle.

Techniques Used: Cell Cycle Assay, Flow Cytometry, Cytometry, Staining

P N elevates Apoptosis and DNA damage: (A) Representative dot-plot of flow-cytometric analysis of P N treated, vehicle treated and D X treated HeLa cells labeled with PI and analyzed by FCS Express.v5 software. (B) Representative dot-plot of flow-cytometric analysis of P N treated, vehicle treated and D X treated HeLa cells labeled with AnnexinV-FITC and analyzed by FCS Express.v5 software. (C) Changes in mean fluorescence intensity (PI) after P N treatment in HeLa cells. (D) Changes in mean fluorescence intensity (AnnexinV-FITC) after P N treatment in HeLa cells. Ordinary one-way ANOVA (Dunnett's multiple comparisons test) was performed to calculate the statistical difference ( p ≤0.05) among all treated groups as compared to vehicle treated group. (E) Changes in apoptotic unit (%) were measured via tunnel assay on incubating P N treated HeLa cells with Caspase 3 inhibitor (Z-DEVD-FMK-Caspase-3. Student t-test (two-tailed, paired) was performed to calculate the statistical difference ( p≤0 . 002 ) among all treated groups as compared to vehicle treated group. P N: Pinostrobin, D X: Doxorubicin, VC: Vehicle control. *, p≤ 0.05; **, p≤ 0.01; ***, p≤ 0.005; ****, p≤ 0.001.
Figure Legend Snippet: P N elevates Apoptosis and DNA damage: (A) Representative dot-plot of flow-cytometric analysis of P N treated, vehicle treated and D X treated HeLa cells labeled with PI and analyzed by FCS Express.v5 software. (B) Representative dot-plot of flow-cytometric analysis of P N treated, vehicle treated and D X treated HeLa cells labeled with AnnexinV-FITC and analyzed by FCS Express.v5 software. (C) Changes in mean fluorescence intensity (PI) after P N treatment in HeLa cells. (D) Changes in mean fluorescence intensity (AnnexinV-FITC) after P N treatment in HeLa cells. Ordinary one-way ANOVA (Dunnett's multiple comparisons test) was performed to calculate the statistical difference ( p ≤0.05) among all treated groups as compared to vehicle treated group. (E) Changes in apoptotic unit (%) were measured via tunnel assay on incubating P N treated HeLa cells with Caspase 3 inhibitor (Z-DEVD-FMK-Caspase-3. Student t-test (two-tailed, paired) was performed to calculate the statistical difference ( p≤0 . 002 ) among all treated groups as compared to vehicle treated group. P N: Pinostrobin, D X: Doxorubicin, VC: Vehicle control. *, p≤ 0.05; **, p≤ 0.01; ***, p≤ 0.005; ****, p≤ 0.001.

Techniques Used: Flow Cytometry, Labeling, Software, Fluorescence, Two Tailed Test

Related Articles

Single Cell Gel Electrophoresis:

Article Title: Non‐canonical cMet regulation by vimentin mediates Plk1 inhibitor–induced apoptosis
Article Snippet: Data were acquired using a flow cytometer (Gallios; Beckman Coulter, Brea, CA) and analyzed using Kaluza software (Beckman Coulter, Brea, CA). .. Comet assay The comet assay was done to detect DNA fragmentation, according to the manufacturer's instructions (Trevigen, Gaithersburg, MD) and as we previously described (Peng et al , ; Sen et al , ). .. After fixation, lysis and electrophoresis slides were stained with Vista Green for fluorescence imaging.

Co-Culture Assay:

Article Title: Agonist redirected checkpoint, PD1-Fc-OX40L, for cancer immunotherapy
Article Snippet: TUNEL assayThe co-culture setup for this assay was identical to the CD107a assay above. .. 1–3 days after co-culture in 96-well plates, DNA fragmentation was assessed using the TiterTACS Colorimetric Apoptosis Detection (TUNEL) Kit from Trevigen, according to manufacturer recommendations. ..

TUNEL Assay:

Article Title: Agonist redirected checkpoint, PD1-Fc-OX40L, for cancer immunotherapy
Article Snippet: TUNEL assayThe co-culture setup for this assay was identical to the CD107a assay above. .. 1–3 days after co-culture in 96-well plates, DNA fragmentation was assessed using the TiterTACS Colorimetric Apoptosis Detection (TUNEL) Kit from Trevigen, according to manufacturer recommendations. ..

Article Title: Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
Article Snippet: HIF-1α – dependent Genes Expression and Determination of Cardiac Cell Apoptosis Total RNA was isolated from whole adult heart in order to measure by Real-Time PCR the expression level of HIF-1α-dependent gene. .. In order to detect cardiac cell apoptosis in the whole heart, three independent methods were used: detection of cytosolic oligonucleosome-bound DNA by ELISA, quantitation of caspase-3 activity by immunoblotting and in situ detection of DNA fragmentation by TUNEL assay (CardioTACS, Trevigen). ..

Article Title: Inhibition of pathological brain angiogenesis through systemic delivery of AAV vector expressing soluble FLT1
Article Snippet: Brain tissue was homogenized in a tissue lysis buffer (Tris buffed saline, proteases inhibitors, 0.1% NP-40). sFLT1 protein was quantified by ELISA assays using Human FLT1 Quantikine ELISA kit (R & D systems Inc., Minneapolis, MN) according to the manufacturer’s instructions. .. TUNEL assay Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was done to identify the extent of DNA fragmentation, using the NeuroTACS II kit (Trevigen, Gaithersburg, MD). ..

Article Title: M1 Muscarinic Receptor Deficiency Attenuates Azoxymethane-Induced Chronic Liver Injury in Mice
Article Snippet: .. To quantify hepatic stellate cell apoptosis, dual staining was performed using a primary antibody to α-SMA followed by terminal UDP-nick end labelling to assess DNA fragmentation (TUNEL; Trevigen, Gaithersburg, MD). ..

Marker:

Article Title: Co-occurrence of opisthorchiasis and diabetes exacerbates morbidity of the hepatobiliary tract disease
Article Snippet: Periductal fibrosis was graded under bright-field microscope into five stages: grade 0 = no fibrosis; grade 1 = mild fibrous expansion of some portal areas; grade 2 = moderate fibrous expansion of most portal areas with short fibrous septa; grade 3 = severe fibrous expansion of most portal areas with occasional portal to portal bridging; and grade 4 = more severe fibrous expansion of most portal areas with marked bridging [ ]. .. DNA fragmentation, an apoptotic marker, within cells fixed in the thin sections of liver was detected in situ (TACS•XL DAB In Situ Apoptosis Detection Kit, Trevigen Inc., Gaithersburg, MD). .. Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL)-positive cells were counted under high power microscopy on 10 non-overlapping areas in each case and are presented as percent positive cells per total number of cells counted in the field [ ].

In Situ:

Article Title: Co-occurrence of opisthorchiasis and diabetes exacerbates morbidity of the hepatobiliary tract disease
Article Snippet: Periductal fibrosis was graded under bright-field microscope into five stages: grade 0 = no fibrosis; grade 1 = mild fibrous expansion of some portal areas; grade 2 = moderate fibrous expansion of most portal areas with short fibrous septa; grade 3 = severe fibrous expansion of most portal areas with occasional portal to portal bridging; and grade 4 = more severe fibrous expansion of most portal areas with marked bridging [ ]. .. DNA fragmentation, an apoptotic marker, within cells fixed in the thin sections of liver was detected in situ (TACS•XL DAB In Situ Apoptosis Detection Kit, Trevigen Inc., Gaithersburg, MD). .. Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL)-positive cells were counted under high power microscopy on 10 non-overlapping areas in each case and are presented as percent positive cells per total number of cells counted in the field [ ].

Article Title: Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
Article Snippet: HIF-1α – dependent Genes Expression and Determination of Cardiac Cell Apoptosis Total RNA was isolated from whole adult heart in order to measure by Real-Time PCR the expression level of HIF-1α-dependent gene. .. In order to detect cardiac cell apoptosis in the whole heart, three independent methods were used: detection of cytosolic oligonucleosome-bound DNA by ELISA, quantitation of caspase-3 activity by immunoblotting and in situ detection of DNA fragmentation by TUNEL assay (CardioTACS, Trevigen). ..

Enzyme-linked Immunosorbent Assay:

Article Title: Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
Article Snippet: HIF-1α – dependent Genes Expression and Determination of Cardiac Cell Apoptosis Total RNA was isolated from whole adult heart in order to measure by Real-Time PCR the expression level of HIF-1α-dependent gene. .. In order to detect cardiac cell apoptosis in the whole heart, three independent methods were used: detection of cytosolic oligonucleosome-bound DNA by ELISA, quantitation of caspase-3 activity by immunoblotting and in situ detection of DNA fragmentation by TUNEL assay (CardioTACS, Trevigen). ..

Quantitation Assay:

Article Title: Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
Article Snippet: HIF-1α – dependent Genes Expression and Determination of Cardiac Cell Apoptosis Total RNA was isolated from whole adult heart in order to measure by Real-Time PCR the expression level of HIF-1α-dependent gene. .. In order to detect cardiac cell apoptosis in the whole heart, three independent methods were used: detection of cytosolic oligonucleosome-bound DNA by ELISA, quantitation of caspase-3 activity by immunoblotting and in situ detection of DNA fragmentation by TUNEL assay (CardioTACS, Trevigen). ..

Activity Assay:

Article Title: Hypoxia/Reoxygenation Cardiac Injury and Regeneration in Zebrafish Adult Heart
Article Snippet: HIF-1α – dependent Genes Expression and Determination of Cardiac Cell Apoptosis Total RNA was isolated from whole adult heart in order to measure by Real-Time PCR the expression level of HIF-1α-dependent gene. .. In order to detect cardiac cell apoptosis in the whole heart, three independent methods were used: detection of cytosolic oligonucleosome-bound DNA by ELISA, quantitation of caspase-3 activity by immunoblotting and in situ detection of DNA fragmentation by TUNEL assay (CardioTACS, Trevigen). ..

End Labeling:

Article Title: Inhibition of pathological brain angiogenesis through systemic delivery of AAV vector expressing soluble FLT1
Article Snippet: Brain tissue was homogenized in a tissue lysis buffer (Tris buffed saline, proteases inhibitors, 0.1% NP-40). sFLT1 protein was quantified by ELISA assays using Human FLT1 Quantikine ELISA kit (R & D systems Inc., Minneapolis, MN) according to the manufacturer’s instructions. .. TUNEL assay Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was done to identify the extent of DNA fragmentation, using the NeuroTACS II kit (Trevigen, Gaithersburg, MD). ..

Staining:

Article Title: M1 Muscarinic Receptor Deficiency Attenuates Azoxymethane-Induced Chronic Liver Injury in Mice
Article Snippet: .. To quantify hepatic stellate cell apoptosis, dual staining was performed using a primary antibody to α-SMA followed by terminal UDP-nick end labelling to assess DNA fragmentation (TUNEL; Trevigen, Gaithersburg, MD). ..

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    Trevigen dna fragmentation
    Cardiomyocyte <t>apoptosis</t> was not detected in 12-week-old FasL Tg hearts by TUNEL or <t>DNA</t> ladder analysis. Paraffin sections of NTg ( a ) and line 61 FasL Tg ( b ) ventricle were stained for apoptotic cells through the labeling of nuclear DNA fragments. The inset in a displays DNase-treated NTg myocardium as a positive control for the TUNEL stain. TUNEL analysis demonstrates no apoptotic or necrotic cardiomyocytes in either NTg or Tg myocardium. Small numbers of TUNEL-positive leukocyte nuclei (arrows) are seen in the interstitium of Tg hearts. The scattered leukocytes, predominantly neutrophils and lymphocytes, are presumably apoptotic. DNA laddering was not detected in NTg, line 70, or line 61 FasL Tg hearts ( c ). Staurosporine-treated rat cardiomyocytes served as positive control for DNA ladder analysis.
    Dna Fragmentation, supplied by Trevigen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Trevigen apoptotic cell system
    Overexpression of RNase L from NL4-3 recombinant proviral DNA (NL4-3/sRL) accelerates HIV-induced cell death. Jurkat cells were transfected with pNL4-3 (○), pNL4-3/Δ nef (◊), pNL4-3/aRL (▴), or pNL4-3/sRL (•) or were mock transfected with salmon sperm DNA (□), and the percentage of <t>apoptotic</t> cells as a function of time was determined by in situ detection of DNA fragmentation. d, days.
    Apoptotic Cell System, supplied by Trevigen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    86
    Trevigen in situ dna fragmentation
    Estrogen-induced osteoclast apoptosis. Osteoclasts were cultured on dentine slices with 0.1 nM E 2 for 24 h. ( A ) A fluorescence micrograph shows normal and <t>apoptotic</t> osteoclasts ( arrowhead ) attached to a dentine slice in a 24-h culture. ×250. ( B ) The TUNEL assay indicates <t>DNA</t> fragmentation in an apoptotic osteoclast. ×250. ( C ) This micrograph, obtained by electron transmission microscopy, demonstrates the gross morphological changes in an apoptotic osteoclast. Nuclear fragments are indicated by the arrowheads. ×2,000.
    In Situ Dna Fragmentation, supplied by Trevigen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Trevigen tacs annexin v apoptosis detection kit
    Activation of MybEnER in clonal populations of M1 and RI-4-11 cells leads to <t>apoptosis</t> and cell cycle arrest. (A) Increase in apoptotic cells as determined by the <t>TACS</t> <t>annexin</t> V apoptosis assay. The y axis depicts the fold increase in the amount of apoptotic cells, and the x axis depicts the hours after stimulation with 4-OHT. Data shown are the mean of three independent results with a standard deviation smaller than 15%. (B) A DNA fragmentation assay used to detect late-stage programmed cell death. DNA laddering is prominent in MybEnER clones following activation of the fusion protein. (C) Flow microfluorometric analysis of the cell cycle distribution in RI-4-11/MybEnER or RI-4-11/EnER cells stimulated with 4-OHT for 24 h. Cell cycle distribution was examined using a Becton Dickinson FACScan and analyzed with the ModFitLT V2.0 program. The proportions of cells in G 0 -G 1 , G 2 -M, and S are shown in boxes in each graph as percentages of the total population.
    Tacs Annexin V Apoptosis Detection Kit, supplied by Trevigen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Cardiomyocyte apoptosis was not detected in 12-week-old FasL Tg hearts by TUNEL or DNA ladder analysis. Paraffin sections of NTg ( a ) and line 61 FasL Tg ( b ) ventricle were stained for apoptotic cells through the labeling of nuclear DNA fragments. The inset in a displays DNase-treated NTg myocardium as a positive control for the TUNEL stain. TUNEL analysis demonstrates no apoptotic or necrotic cardiomyocytes in either NTg or Tg myocardium. Small numbers of TUNEL-positive leukocyte nuclei (arrows) are seen in the interstitium of Tg hearts. The scattered leukocytes, predominantly neutrophils and lymphocytes, are presumably apoptotic. DNA laddering was not detected in NTg, line 70, or line 61 FasL Tg hearts ( c ). Staurosporine-treated rat cardiomyocytes served as positive control for DNA ladder analysis.

    Journal: Journal of Clinical Investigation

    Article Title: Proinflammatory consequences of transgenic Fas ligand expression in the heart

    doi:

    Figure Lengend Snippet: Cardiomyocyte apoptosis was not detected in 12-week-old FasL Tg hearts by TUNEL or DNA ladder analysis. Paraffin sections of NTg ( a ) and line 61 FasL Tg ( b ) ventricle were stained for apoptotic cells through the labeling of nuclear DNA fragments. The inset in a displays DNase-treated NTg myocardium as a positive control for the TUNEL stain. TUNEL analysis demonstrates no apoptotic or necrotic cardiomyocytes in either NTg or Tg myocardium. Small numbers of TUNEL-positive leukocyte nuclei (arrows) are seen in the interstitium of Tg hearts. The scattered leukocytes, predominantly neutrophils and lymphocytes, are presumably apoptotic. DNA laddering was not detected in NTg, line 70, or line 61 FasL Tg hearts ( c ). Staurosporine-treated rat cardiomyocytes served as positive control for DNA ladder analysis.

    Article Snippet: Evidence of apoptosis or necrosis was assessed by detection of nuclear DNA fragmentation in paraffin-fixed myocardial sections by terminal deoxynucleotidyl transferase–mediated dUTP nick end-labeling (CardioTACS Blue TUNEL staining; Trevigen Inc., Gaithersburg, Maryland, USA) according to the manufacturer’s instructions.

    Techniques: TUNEL Assay, Staining, Labeling, Positive Control, DNA Laddering

    Overexpression of RNase L from NL4-3 recombinant proviral DNA (NL4-3/sRL) accelerates HIV-induced cell death. Jurkat cells were transfected with pNL4-3 (○), pNL4-3/Δ nef (◊), pNL4-3/aRL (▴), or pNL4-3/sRL (•) or were mock transfected with salmon sperm DNA (□), and the percentage of apoptotic cells as a function of time was determined by in situ detection of DNA fragmentation. d, days.

    Journal: Journal of Virology

    Article Title: Regulation of Human Immunodeficiency Virus Replication by 2?,5?-Oligoadenylate-Dependent RNase L

    doi:

    Figure Lengend Snippet: Overexpression of RNase L from NL4-3 recombinant proviral DNA (NL4-3/sRL) accelerates HIV-induced cell death. Jurkat cells were transfected with pNL4-3 (○), pNL4-3/Δ nef (◊), pNL4-3/aRL (▴), or pNL4-3/sRL (•) or were mock transfected with salmon sperm DNA (□), and the percentage of apoptotic cells as a function of time was determined by in situ detection of DNA fragmentation. d, days.

    Article Snippet: DNA fragmentation in situ was measured with the Trevigen Apoptotic Cell System by using terminal deoxynucleotide transferase (15 U) in 0.05 M Tris (pH 7.5)–5 mM MgCl2 –0.6 mM β-mercaptoethanesulfonic acid–50 μg of bovine serum albumin–0.25 mM biotinylated nucleotides (dNTPs) at 37°C for 60 min.

    Techniques: Over Expression, Recombinant, Transfection, In Situ

    Estrogen-induced osteoclast apoptosis. Osteoclasts were cultured on dentine slices with 0.1 nM E 2 for 24 h. ( A ) A fluorescence micrograph shows normal and apoptotic osteoclasts ( arrowhead ) attached to a dentine slice in a 24-h culture. ×250. ( B ) The TUNEL assay indicates DNA fragmentation in an apoptotic osteoclast. ×250. ( C ) This micrograph, obtained by electron transmission microscopy, demonstrates the gross morphological changes in an apoptotic osteoclast. Nuclear fragments are indicated by the arrowheads. ×2,000.

    Journal: The Journal of Experimental Medicine

    Article Title: Estrogen Inhibits Bone Resorption by Directly Inducing Apoptosis of the Bone-resorbing Osteoclasts

    doi:

    Figure Lengend Snippet: Estrogen-induced osteoclast apoptosis. Osteoclasts were cultured on dentine slices with 0.1 nM E 2 for 24 h. ( A ) A fluorescence micrograph shows normal and apoptotic osteoclasts ( arrowhead ) attached to a dentine slice in a 24-h culture. ×250. ( B ) The TUNEL assay indicates DNA fragmentation in an apoptotic osteoclast. ×250. ( C ) This micrograph, obtained by electron transmission microscopy, demonstrates the gross morphological changes in an apoptotic osteoclast. Nuclear fragments are indicated by the arrowheads. ×2,000.

    Article Snippet: To detect in situ DNA fragmentation of apoptotic osteoclasts, we employed the TUNEL (TdT-mediated dUTP-biotin nick end-labeling) assay by using a TACS Blue Label™ kit (Trevigen, Inc., Gaithersburg, MD) according to the procedure recommended by the manufacturer.

    Techniques: Cell Culture, Fluorescence, TUNEL Assay, Transmission Assay, Microscopy

    Activation of MybEnER in clonal populations of M1 and RI-4-11 cells leads to apoptosis and cell cycle arrest. (A) Increase in apoptotic cells as determined by the TACS annexin V apoptosis assay. The y axis depicts the fold increase in the amount of apoptotic cells, and the x axis depicts the hours after stimulation with 4-OHT. Data shown are the mean of three independent results with a standard deviation smaller than 15%. (B) A DNA fragmentation assay used to detect late-stage programmed cell death. DNA laddering is prominent in MybEnER clones following activation of the fusion protein. (C) Flow microfluorometric analysis of the cell cycle distribution in RI-4-11/MybEnER or RI-4-11/EnER cells stimulated with 4-OHT for 24 h. Cell cycle distribution was examined using a Becton Dickinson FACScan and analyzed with the ModFitLT V2.0 program. The proportions of cells in G 0 -G 1 , G 2 -M, and S are shown in boxes in each graph as percentages of the total population.

    Journal: Molecular and Cellular Biology

    Article Title: Regulation of the Resident Chromosomal Copy of c-myc by c-Myb Is Involved in Myeloid Leukemogenesis

    doi:

    Figure Lengend Snippet: Activation of MybEnER in clonal populations of M1 and RI-4-11 cells leads to apoptosis and cell cycle arrest. (A) Increase in apoptotic cells as determined by the TACS annexin V apoptosis assay. The y axis depicts the fold increase in the amount of apoptotic cells, and the x axis depicts the hours after stimulation with 4-OHT. Data shown are the mean of three independent results with a standard deviation smaller than 15%. (B) A DNA fragmentation assay used to detect late-stage programmed cell death. DNA laddering is prominent in MybEnER clones following activation of the fusion protein. (C) Flow microfluorometric analysis of the cell cycle distribution in RI-4-11/MybEnER or RI-4-11/EnER cells stimulated with 4-OHT for 24 h. Cell cycle distribution was examined using a Becton Dickinson FACScan and analyzed with the ModFitLT V2.0 program. The proportions of cells in G 0 -G 1 , G 2 -M, and S are shown in boxes in each graph as percentages of the total population.

    Article Snippet: Early-stage apoptosis was detected using the TACS annexin V apoptosis detection kit (Trevigen, Inc.) according to the manufacturer's instructions.

    Techniques: Activation Assay, Apoptosis Assay, Standard Deviation, DNA Fragmentation Assay, DNA Laddering, Clone Assay, Flow Cytometry