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multiparameter apoptosis hitkit  (Thermo Fisher)


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    Thermo Fisher multiparameter apoptosis hitkit
    The effects of TXS inhibition on tumour cell <t>apoptosis</t> . A) Apoptosis was induced in a dose-dependant manner following 24 h treatment with ozagrel, relative to untreated control cells. Cell health following ozagrel treatment was assessed using 3 spectrally distinct flourophores to examine nuclei, f -actin (marker of cytoskeletal integrity), and mitochondrial mass/potential. Reduced f -actin levels demonstrate a loss in cellular integrity during apoptosis. Membrane blebbing also occurs and mitochondrial activity occurs, coupled with a loss of potential across the mitochondrial membrane. These markers were quantified by the Kinetic Scan HCS reader and are represented (B). Similar observations were made in SKMES-1 cells (data not shown). Apoptosis was confirmed following selective TXS inhibition by Cell Death Detection ELISA and DNA laddering in both cell lines (A-549 shown as representative). Cell Death ELISA demonstrated increased apoptosis in a concentration dependant manner, with fold induction expressed as a ratio of control cells (C). DNA laddering was also observed following ozagrel treatment at both 500 nM and 5 μM concentrations (D).
    Multiparameter Apoptosis Hitkit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/hitkit/pmc03074522-200-1-5
    Average 90 stars, based on 1 article reviews
    multiparameter apoptosis hitkit - by Bioz Stars, 2026-09
    90/100 stars

    Images

    1) Product Images from "Examination of thromboxane synthase as a prognostic factor and therapeutic target in non-small cell lung cancer"

    Article Title: Examination of thromboxane synthase as a prognostic factor and therapeutic target in non-small cell lung cancer

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-10-25

    The effects of TXS inhibition on tumour cell apoptosis . A) Apoptosis was induced in a dose-dependant manner following 24 h treatment with ozagrel, relative to untreated control cells. Cell health following ozagrel treatment was assessed using 3 spectrally distinct flourophores to examine nuclei, f -actin (marker of cytoskeletal integrity), and mitochondrial mass/potential. Reduced f -actin levels demonstrate a loss in cellular integrity during apoptosis. Membrane blebbing also occurs and mitochondrial activity occurs, coupled with a loss of potential across the mitochondrial membrane. These markers were quantified by the Kinetic Scan HCS reader and are represented (B). Similar observations were made in SKMES-1 cells (data not shown). Apoptosis was confirmed following selective TXS inhibition by Cell Death Detection ELISA and DNA laddering in both cell lines (A-549 shown as representative). Cell Death ELISA demonstrated increased apoptosis in a concentration dependant manner, with fold induction expressed as a ratio of control cells (C). DNA laddering was also observed following ozagrel treatment at both 500 nM and 5 μM concentrations (D).
    Figure Legend Snippet: The effects of TXS inhibition on tumour cell apoptosis . A) Apoptosis was induced in a dose-dependant manner following 24 h treatment with ozagrel, relative to untreated control cells. Cell health following ozagrel treatment was assessed using 3 spectrally distinct flourophores to examine nuclei, f -actin (marker of cytoskeletal integrity), and mitochondrial mass/potential. Reduced f -actin levels demonstrate a loss in cellular integrity during apoptosis. Membrane blebbing also occurs and mitochondrial activity occurs, coupled with a loss of potential across the mitochondrial membrane. These markers were quantified by the Kinetic Scan HCS reader and are represented (B). Similar observations were made in SKMES-1 cells (data not shown). Apoptosis was confirmed following selective TXS inhibition by Cell Death Detection ELISA and DNA laddering in both cell lines (A-549 shown as representative). Cell Death ELISA demonstrated increased apoptosis in a concentration dependant manner, with fold induction expressed as a ratio of control cells (C). DNA laddering was also observed following ozagrel treatment at both 500 nM and 5 μM concentrations (D).

    Techniques Used: Inhibition, Marker, Activity Assay, Enzyme-linked Immunosorbent Assay, DNA Laddering, Concentration Assay

    Effect of stable TXS -over-expression on tumour cell survival . Apoptosis was measured in TXS stable transfectants, and corresponding controls (wild-type and empty vector) following 48 h (A) and 72 h (B) serum-starvation by flow cytometry. Representative dot plots following 72 h serum starvation are shown for wild-type (C), empty vector (D) and TXS overexpressing (E) cells. Graphical data is represented at a percentage of the empty vector control, which was set to 100%. Data is expressed as mean ± SEM. n=3 independent experiments.
    Figure Legend Snippet: Effect of stable TXS -over-expression on tumour cell survival . Apoptosis was measured in TXS stable transfectants, and corresponding controls (wild-type and empty vector) following 48 h (A) and 72 h (B) serum-starvation by flow cytometry. Representative dot plots following 72 h serum starvation are shown for wild-type (C), empty vector (D) and TXS overexpressing (E) cells. Graphical data is represented at a percentage of the empty vector control, which was set to 100%. Data is expressed as mean ± SEM. n=3 independent experiments.

    Techniques Used: Over Expression, Plasmid Preparation, Flow Cytometry

    Related Articles

    Staining:

    Article Title: Specific modulation of astrocyte inflammation by inhibition of mixed lineage kinases with CEP-1347.
    Article Snippet: .. Primary astrocytes were stimulated in 96-well plates and the Cellomics HitKit was used according to the manufacturer’s protocol to stain the cells. ..

    Article Title: Specific modulation of astrocyte inflammation by inhibition of mixed lineage kinases with CEP-1347.
    Article Snippet: .. Primary astrocytes were stimulated in 96-well plates and the Cellomics HitKit (Reading, U.K.) was used according to the manufacturer’s protocol to stain the cells. ..

    Activation Assay:

    Article Title: Activation of stress-related signalling pathway in human cells upon SiO 2 nanoparticles exposure as an early indicator of cytotoxicity
    Article Snippet: The rate of cell viability and proliferation were assessed by the automated analysis of the nuclear count and morphology (DAPI filter); in parallel to the fluorescent staining intensities reflecting cell permeability (FITC filter) and lysosomal mass/pH changes (TRITC filter) were also quantified for each individual cell present in the examined microscopic fields by IN Cell Investigator (GE Healthcare, UK). .. ATF-2 activation was measured using Cellomics HitKit ® as per manufacturer's instructions (Thermo Fisher Scientific Inc., USA). ..

    Article Title: Activation of stress-related signalling pathway in human cells upon SiO 2 nanoparticles exposure as an early indicator of cytotoxicity
    Article Snippet: Dulbecco's modified Eagle medium (DMEM), RPMI 1640 and foetal bovine serum were from Gibco (Invitrogen, BioSciences Ltd., Dublin, Ireland). .. HitKitTM for ATF-2 activation and multiparametric cytotoxicity assay 1 (MPCT1) were from Thermo Fisher Scientific (Thermo Fisher Scientific Inc., USA). .. Rabbit monoclonal anti-JNK, anti-phospho-JNK, anti-p38, anti-phospho- p38 and horseradish peroxidase conjugated anti-rabbit antibodies were from Cell Signaling Technology (Danvers, MA, USA).

    Cytotoxicity Assay:

    Article Title: Activation of stress-related signalling pathway in human cells upon SiO 2 nanoparticles exposure as an early indicator of cytotoxicity
    Article Snippet: Dulbecco's modified Eagle medium (DMEM), RPMI 1640 and foetal bovine serum were from Gibco (Invitrogen, BioSciences Ltd., Dublin, Ireland). .. HitKitTM for ATF-2 activation and multiparametric cytotoxicity assay 1 (MPCT1) were from Thermo Fisher Scientific (Thermo Fisher Scientific Inc., USA). .. Rabbit monoclonal anti-JNK, anti-phospho-JNK, anti-p38, anti-phospho- p38 and horseradish peroxidase conjugated anti-rabbit antibodies were from Cell Signaling Technology (Danvers, MA, USA).



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    The effects of TXS inhibition on tumour cell <t>apoptosis</t> . A) Apoptosis was induced in a dose-dependant manner following 24 h treatment with ozagrel, relative to untreated control cells. Cell health following ozagrel treatment was assessed using 3 spectrally distinct flourophores to examine nuclei, f -actin (marker of cytoskeletal integrity), and mitochondrial mass/potential. Reduced f -actin levels demonstrate a loss in cellular integrity during apoptosis. Membrane blebbing also occurs and mitochondrial activity occurs, coupled with a loss of potential across the mitochondrial membrane. These markers were quantified by the Kinetic Scan HCS reader and are represented (B). Similar observations were made in SKMES-1 cells (data not shown). Apoptosis was confirmed following selective TXS inhibition by Cell Death Detection ELISA and DNA laddering in both cell lines (A-549 shown as representative). Cell Death ELISA demonstrated increased apoptosis in a concentration dependant manner, with fold induction expressed as a ratio of control cells (C). DNA laddering was also observed following ozagrel treatment at both 500 nM and 5 μM concentrations (D).
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    Image Search Results


    The effects of TXS inhibition on tumour cell apoptosis . A) Apoptosis was induced in a dose-dependant manner following 24 h treatment with ozagrel, relative to untreated control cells. Cell health following ozagrel treatment was assessed using 3 spectrally distinct flourophores to examine nuclei, f -actin (marker of cytoskeletal integrity), and mitochondrial mass/potential. Reduced f -actin levels demonstrate a loss in cellular integrity during apoptosis. Membrane blebbing also occurs and mitochondrial activity occurs, coupled with a loss of potential across the mitochondrial membrane. These markers were quantified by the Kinetic Scan HCS reader and are represented (B). Similar observations were made in SKMES-1 cells (data not shown). Apoptosis was confirmed following selective TXS inhibition by Cell Death Detection ELISA and DNA laddering in both cell lines (A-549 shown as representative). Cell Death ELISA demonstrated increased apoptosis in a concentration dependant manner, with fold induction expressed as a ratio of control cells (C). DNA laddering was also observed following ozagrel treatment at both 500 nM and 5 μM concentrations (D).

    Journal: Molecular Cancer

    Article Title: Examination of thromboxane synthase as a prognostic factor and therapeutic target in non-small cell lung cancer

    doi: 10.1186/1476-4598-10-25

    Figure Lengend Snippet: The effects of TXS inhibition on tumour cell apoptosis . A) Apoptosis was induced in a dose-dependant manner following 24 h treatment with ozagrel, relative to untreated control cells. Cell health following ozagrel treatment was assessed using 3 spectrally distinct flourophores to examine nuclei, f -actin (marker of cytoskeletal integrity), and mitochondrial mass/potential. Reduced f -actin levels demonstrate a loss in cellular integrity during apoptosis. Membrane blebbing also occurs and mitochondrial activity occurs, coupled with a loss of potential across the mitochondrial membrane. These markers were quantified by the Kinetic Scan HCS reader and are represented (B). Similar observations were made in SKMES-1 cells (data not shown). Apoptosis was confirmed following selective TXS inhibition by Cell Death Detection ELISA and DNA laddering in both cell lines (A-549 shown as representative). Cell Death ELISA demonstrated increased apoptosis in a concentration dependant manner, with fold induction expressed as a ratio of control cells (C). DNA laddering was also observed following ozagrel treatment at both 500 nM and 5 μM concentrations (D).

    Article Snippet: The Multiparamater Apoptosis HitKit from Cellomics provides High Content Screening qualified fluorescent reagents for simultaneous measurement of 3 fundamental parameters of apoptosis.

    Techniques: Inhibition, Marker, Activity Assay, Enzyme-linked Immunosorbent Assay, DNA Laddering, Concentration Assay

    Effect of stable TXS -over-expression on tumour cell survival . Apoptosis was measured in TXS stable transfectants, and corresponding controls (wild-type and empty vector) following 48 h (A) and 72 h (B) serum-starvation by flow cytometry. Representative dot plots following 72 h serum starvation are shown for wild-type (C), empty vector (D) and TXS overexpressing (E) cells. Graphical data is represented at a percentage of the empty vector control, which was set to 100%. Data is expressed as mean ± SEM. n=3 independent experiments.

    Journal: Molecular Cancer

    Article Title: Examination of thromboxane synthase as a prognostic factor and therapeutic target in non-small cell lung cancer

    doi: 10.1186/1476-4598-10-25

    Figure Lengend Snippet: Effect of stable TXS -over-expression on tumour cell survival . Apoptosis was measured in TXS stable transfectants, and corresponding controls (wild-type and empty vector) following 48 h (A) and 72 h (B) serum-starvation by flow cytometry. Representative dot plots following 72 h serum starvation are shown for wild-type (C), empty vector (D) and TXS overexpressing (E) cells. Graphical data is represented at a percentage of the empty vector control, which was set to 100%. Data is expressed as mean ± SEM. n=3 independent experiments.

    Article Snippet: The Multiparamater Apoptosis HitKit from Cellomics provides High Content Screening qualified fluorescent reagents for simultaneous measurement of 3 fundamental parameters of apoptosis.

    Techniques: Over Expression, Plasmid Preparation, Flow Cytometry