To this end, we selected 14 well-known genotoxic compounds and compared them with 10 nongenotoxic chemicals, using CHO-9 cells because they are well characterized as to DNA repair and DDR. We quantified gamma-H2AX foci manually and automatically. In addition, total gamma-H2AX activation was determined by flow cytometry.

6 gamma-H2AX Primary Antibodies: Thermo Fisher antibodies are validated for applications including western blotting, immunocytochemistry, flow cytometry, and chromatin immunoprecipitation.

Prepare 70% Validation of a flow cytometry-based detection of γ-H2AX, to measure DNA damage for clinical applications. Artikel i vetenskaplig tidskrift, refereegranskad. A control for the day-to-day normalization of the flow cytometry γ-H2AX assay for clinical routine. Artikel i vetenskaplig tidskrift, refereegranskad. Författare.

Lastly, flow cytometry has been used to analyze γ-H2AX; however, flow cytometry methods are not readily integrated into HTS. Although each of the aforementioned methods of evaluating γ-H2AX is effective and has provided important information, there is still a need for an analytical high throughput assay that is capable of screening radiomodifying drugs across diverse cell lines and in vivo 2011-09-23 · Here, measurements of γ-H2AX immunofluorescence by microscopy and flow cytometry were compared as rapid biodosimetric tools for whole and partial body exposures. Ex vivo uniformly X-irradiated blood lymphocytes from one donor were used to generate a universal biexponential calibration function for γ-H2AX foci/intensity yields per unit dose for time points up to 96 hours post exposure. 2010-02-04 · Gamma-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic Acids Res 2008; 36 : 5678–5694. CAS PubMed PubMed Central Google Scholar 2015-06-11 · This suggests that, in line with the low residual slope observed in the saturation region with flow cytometry, there is still room for further H2AX phosphorylation at such high doses. These results are further supported by the direct comparison of flow cytometry data and integral fluorescence intensity as extracted from microscopy pictures which is shown in S1 Fig .

2019-08-22 · The objective of the present study was to develop a rapid, high-throughput γ-H2AX assay based on imaging flow cytometry (IFC) using the ImageStream® X Mk II (ISX) platform to evaluate DNA double strand break (DSB) repair kinetics in human peripheral blood cells after exposure to ionizing irradiation.

Histone H2A.X (H2AX) is a member of the histone H2A family which is one of the four core histones making up the nucleosome core particle. In eukaryotes, DNA double strand breaks (DSBs) have been shown to trigger the phosphorylation of serine 139 at the carboxy terminus of histone H2AX resulting in gamma-H2AX. This process is believed to play a key role in the repair of DNA damage.

2011-09-23 · Here, measurements of γ-H2AX immunofluorescence by microscopy and flow cytometry were compared as rapid biodosimetric tools for whole and partial body exposures. Ex vivo uniformly X-irradiated blood lymphocytes from one donor were used to generate a universal biexponential calibration function for γ-H2AX foci/intensity yields per unit dose for time points up to 96 hours post exposure.

Vi fann att fenotiaziner delade förmågan att fördröja γ H2AX upplösning i to cellcycle positions was determined by two-color flow cytometry using DAPI as the Immunohistokemisk färgning; Biochemistry assays; Flow cytometry analysis SOD1 (Abcam), Sirt1 (Abcam), γ-H2AX (Ser139) (Cell Signaling Technology), DNA damage can be studied by flow cytometric analysis of gamma-H2AX in human peripheral lymphocytes. The exposure to multiple CT scans causes more double strand breaks as compared to single scan. DNA damage can be studied by flow cytometric analysis of gamma-H2AX in human peripheral lymphocytes.

Although flow cytometry has been used to quantify DNA damage, it is still unexplored when it comes to low dose radiation and its effects,. This process is believed to play a key role in the repair of DNA damage. In this study, we established a flow cytometry (FCM) system for measuring radiation-induced phosphorylated histone H2AX (gammaH2AX) in cultured human T lymphocytes to evaluate individual radiation sensitivity in vitro. At present, flow cytometry is the most rapid method for detection of DSBs and cell viability. In this chapter, we provide our experience and methodological modification of flow cytometry protocol for the detection of γ-H2AX, a well-known marker of DSBs, in fixed mammalian fibroblasts. The objective of the present study was to develop a rapid, high-throughput γ-H2AX assay based on imaging flow cytometry (IFC) using the ImageStream® X Mk II (ISX) platform to evaluate DNA double strand break (DSB) repair kinetics in human peripheral blood cells after exposure to ionizing irradiation.
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2019-08-22 · The objective of the present study was to develop a rapid, high-throughput γ-H2AX assay based on imaging flow cytometry (IFC) using the ImageStream® X Mk II (ISX) platform to evaluate DNA double strand break (DSB) repair kinetics in human peripheral blood cells after exposure to ionizing irradiation.

However, γ-H2AX is not able to monitor repair kinetics within the first 60 min postirradiation, a period when most DSBs undergo repair. Öppna fliken parametrar i fönstret cytometer och välj parametrarna enligt tabell 1. Vi tackar Flow Cytometry Facility team på tyska cancer Research Center validation for radiation-induced gamma-H2AX foci in human cells. Att kvantifiera procentandelen av celler som har γ-H2AX-positiva foci, räkna varje cell Att kvantifiera antalet av γ-H2AX foci, räkna γ-H2AX foci per cell.
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H2AX, H2a.x, H2afx, H2A/X 89 citations have been found for this product All applications Flow cytometry/Cell sorting (FC/FACS) Immunocytochemistry (ICC) Immunocytochemistry-immunoflourescence (ICC-IF) Immunofluorescence (IF) Immunohistochemistry (IHC) Immunohistochemistry-immunofluorescence (IHC-IF) Immunohistochemistry-paraffin (IHC-P) Immunoprecipitation (IP) Western Blotting (WB)

After treatment, 1 mL of 0.1% BSA‐PBS was added to the samples and PBMCs were pelleted (5 min at 2000 g) followed by fixation in 0.25% paraformaldehyde‐PBS (8 × 10 6 cells/mL), for 10 min on ice. At present, flow cytometry is the most rapid method for detection of DSBs and cell viability. In this chapter, we provide our experience and methodological modification of flow cytometry protocol Measurement of c-H2AX by Flow Cytometry H2AX phosphorylation was analyzed by ﬂow cyto-metry analysis as previously described (23), with small modiﬁcations.