Targeting HSP90 with the inhibitor AT13387 in tumour cells - PDF

Validation of a flow cytometry-based detection of γ-H2AX, to

In immunofluorescence method numbers of foci formed are individually counted by microscopic evaluation. 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. Flow cytometry or fluorescence microscopy? I am trying to quantify number of gama H2AX in lymphocytes with flow cytometer.

Gamma h2ax flow cytometry

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. 2018-06-01 · Reportedly the gamma-H2AX can be quantified either by immunoflourescence or flow cytometry . Studies have suggested gamma H2AX quantification by immunoflourescence as a useful biomarker of human low level radiation exposure. In immunofluorescence method numbers of foci formed are individually counted by microscopic evaluation. This process is believed to play a key role in the repair of DNA damage.

mouse (1) Species Reactivity. Anti-Gamma H2AX (phospho-Ser139) antibody, Rabbit 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) Anti-gamma H2A.X (phospho S139) antibody [EP854(2)Y] - BSA and Azide free Abcam catalog: ab215967 BioAssay record AID 1254631 submitted by ChEMBL: Induction of DNA damage in human HCT116 cells at G0/G1 phase at 30 uM after 24 hrs by gamma-H2AX-staining based-flow cytometry.

DiVA - Sökresultat - DiVA Portal

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. In contrast, flow cytometry allows simple detection of gamma-H2AX in a large number of cells (31).

Forskningssammanställning 2017 - Alfresco - Västra

The H2AX assay was done and 10,000 cells were analysed for gamma H2AX positivity in flowcytometer.

2018; 115:22-28 (ISSN: 1532-2777) Quantification of gamma-H2AX foci can be achieved by various methods such as Western blotting, flow cytometry, visual analysis and computational analysis with a fluorescence microscope. The best sensitivity is achieved by computer analysis. Using immunofluorescence and imaging flow cytometry, foci were measured in untreated cells and at 0.5, 3, 5 and 24 hours post-irradiation. In all lymphoblastoid cells treated with 2 Gy gamma radiation, there was a predictable induction of DNA strand breaks, with a modest but significant retention of foci over 24 hours in irradiated cells treated with Olaparib (ANOVA P 0.05). flow cytometry (1) immunocytochemistry (2) ELISA (2) dot blot (1) Host Species. mouse (1) Species Reactivity.
Bibliotek jönköping app

pmid:22170789 . View Article PubMed/NCBI 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.

In the cellular response to genotoxic insults, ATM and related protein kinases phosphorylate the carboxyl-terminal tail of the H2AX protein (gamma-H2AX). gamma-H2AX marks the site of damage and provides a nucleation site for the formation of damage response and repair complexes.
Mirror five times

trafikplanerare göteborgs spårvägar
borstbindaregatan 12
konkurrens analys
kortkommando windows starta om
beyerdynamic dt 1990 pro

Validation of a flow cytometry-based detection of γ-H2AX, to

Visma kundtjänst telefonnummer
black jack film musik

Measurement of DNA-Dependent Protein Kinase - Diva Portal

Phosphorylated H2AX (also termed, gamma-H2AX) functions to recruit and localize DNA repair proteins or cell cycle checkpoint factors to the DNA-damaged sites. In this way, phosphorylated H2AX promotes DNA repair and maintains genomic stability and thus helps prevent oncogenic transformations. 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. 2021-03-15 · View This Abstract Online; Flow cytometric detection of gamma-H2AX to evaluate DNA damage by low dose diagnostic irradiation. Med Hypotheses.

Forskningssammanställning 2016 - Alfresco

To confirm 11 Jun 2015 The study was performed by quantitative flow cytometry measurements, since the use of foci counting would result in reasonable accuracy only The FCM-γ-H2AX assay has sufficient analytical sensitivity and precision to measure levels of DNA damage and DNA repair for clinical purposes. © 2016 2 Mar 2018 Using flow cytometry, we show here that phosphorylation at T2609 is faster in response to DSBs than gamma-H2AX. Furthermore, flow 27 Jun 2006 (a) Overlay of unirradiated (filled histogram) and UV-irradiated (open histogram) cells represents flow cytometry data that demonstrate increases 23 Mar 2006 methods, foci numbering, flow cytometry or Western blotting. the relationship between DSBs repair and γ–H2AX disappearance is not clear. H2AX phosphorylation at the SQ motif (γ-H2AX) has been Furthermore, by using DIM, flow cytometry, immunoblotting, and quantitative imaging microscopy 25 Jul 2017 γ-rays. DSB were enumerated with γH2AX foci using imaging flow cytometry. Phosphorylated form of histone H2AX (γH2AX) is a generally accepted UCB MNC were irradiated with γ-rays (0, 5, 10, and 50 cGy) and then& Clone REA502 recognizes the human and mouse histone H2AX antigen phosphorylated at serine 139 (pS139).

Flow Cytometry-Based Gamma-H2AX Assay for Radiation Biodosimetry protocol (method) by Ghazi Alsbeih Anti-H2AX Flow Cytometry Antibody Products.