4 Nov 2016 X, Gamma-H2AX; Isotype: Mouse IgG1, κ; Ave. Rating: Submit a X- Phosphorylated (Ser139) Antibody for Flow Cytometry 1. Prepare 70%

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.

The purpose of the present study was to assess immediate DNA damage after exposure to low level of ionizing radiation by the flow cytometric method of gamma-H2AX. Here, measurements of γ-H2AX immunofluorescence by microscopy and flow cytometry were compared as rapid biodosimetric tools for whole and partial body exposures. View This Abstract Online; Flow cytometric detection of gamma-H2AX to evaluate DNA damage by low dose diagnostic irradiation. Med Hypotheses. 2018; 115:22-28 (ISSN: 1532-2777) Gamma H2AX (gammaH2AX) is the phosphorylated version of histone H2AX and is a marker for double-stranded breaks (DSBs) caused by DNA damage (1-4). H2AX is a variant of histone H2A, one of the histone core molecules forming the nucleosome, and is a vital component in repairing DNA damage (1-4). Flow cytometry or fluorescence microscopy?

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

Validated in WB, IHC, ICC, Flow Cyt and tested in Human. (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma; Flow Cytometry - Anti-gamma H2A.

6 gamma-H2AX Primary Antibodies: Thermo Fisher antibodies are validated for applications including western blotting, immunocytochemistry, flow cytometry, and chromatin immunoprecipitation. 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 contrast, flow cytometry allows simple detection of gamma-H2AX in a large number of cells (31). Several reports show that the level of gamma-H2AX as detected by flow cytometry correlates well with the number of DNA strand breaks, to the level of cell death and radiosensitivity (32–34).

At present, flow cytometry is the most rapid method for detection of DSBs and cell viability.

DNA damage can be studied by flow cytometric analysis of gamma-H2AX in human peripheral lymphocytes. H2AX phosphorylation was analyzed by flow cytometry analysis as previously described 23, with small modifications. 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.
Usa tullar kina

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).

Additional reported applications (for the relevant formats of this clone) include: immunohistochemistry on paraffin embedded sections 2, immunofluorescence microscopy 3-9, Western blotting 10-12, and flow cytometry 1,13. Clone 2F3 cross-reacts with mouse 4. Intracellular staining protocol for Anti-H2A.X-Phosphorylated (Ser139) Antibody for Flow Furthermore, flow cytometric analysis of T2609 resulted in a better representation of fast repair kinetics than analysis of gamma-H2AX. In cells with reduced ligase IV activity, and wild-type cells where DNA-dependent protein kinase activity was inhibited, the reduced DSB repair capacity was observed by T2609 evaluation using flow cytometry.
Objektorienterad programmering php

flingan
lunch perrongen markaryd
vad heter distanskurs på engelska
lantbutiken
introduktion till samhällsvetenskaplig analys

The H2AX assay was done and 10,000 cells were analysed for gamma H2AX positivity in flowcytometer. Results Significant gamma-H2AX positivity was found in cases versus control, the most significant DNA damage amongst cases was observed in cases with multiple CT scans.

Balkan folk dance
varför betalar inte företag moms

Flow Cytometry: gamma H2AX [p Ser139] Antibody [NB100-384] - Analysis of gamma-H2AX in EPE Treated Jurkat Cells. Cells were treated for 3 hrs in 5ug/ml etoposide,

Cells were treated for 3 hrs in 5ug/ml etoposide, 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. The H2AX assay was done and 10,000 cells were analysed for gamma H2AX positivity in flowcytometer. Results Significant gamma-H2AX positivity was found in cases versus control, the most significant DNA damage amongst cases was observed in cases with multiple CT scans. The flow cytometry analysis of γH2AX was used in another study to confirm the genotoxic potential of selected compounds in HepG2 cells. The compounds were classified as true genotoxicants, nongenotoxicants and false genotoxicants showing positive results in in vitro genotoxicity assays, but negative results in in vivo genotoxicity assays.