Sektion V - Forskningsoutput - Lunds universitet

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Reactivity Human, Mouse, Rat, Canine Host Rabbit Isotype IgG Vial size 0.1 ml Concentration 1.0 mg 2016-03-31 · gamma-h2ax-phospho-s139-antibody-ab2893.pdf. Send me a copy of this email Flow Cytometry abreview for Anti-gamma H2A.X (phospho S139) antibody Excellent. 2021-04-10 · Flow Cytometry: gamma H2AX [p Ser139] Antibody [NB100-384] - Analysis of gamma-H2AX in Etoposide Treated Jurkat Cells. Cells were treated for 3 hrs in 5ug/ml etoposide, fixed in 1.5% PFA, and permeabilized in 90% Methanol.

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). 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. 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.

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60.00. 70.00. 0.00%. Avg Ped RR (N=8).

Cell Cycle-specific Measurement of γH2AX and Apoptosis

P-H2AX flow cytometry assay in the clinic in a controlled manner. Finally, using immunostaining in optimized method for measurement of gamma-H2AX in.

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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.

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.
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The prognostic value of gamma-H2AX is indicated in certain cancer types, such as breast or endometrial cancer, but further investigation is needed to establish gamma-H2AX as a prognostic marker. 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.

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1 million cells were stained with 0.5 ug anti-KLH or anti-H2AX NB100-384 and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Phosphorylated H2AX (also termed, gamma-H2AX) functions to recruit and localize DNA repair Flow cytometric analysis of H2AX (pS139) expression in. 6 Sep 2006 We applied a flow cytometric method to quantify IR-induced histone H2AX phosphorylation at serine 139 H2AX, at the sites of DSB damage that precedes the invo- IdU-induced DNA double-strand breaks with gamma-. 28 Sep 2018 Subsequently, γH2AX levels were assessed by flow cytometry, rapid phosphorylation of the core histone variant H2AX at serine 139 is 1 Sep 2019 The presented flow cytometric method has been developed to for analysis of foci: validation for radiation-induced gamma-H2AX foci in 26 Jun 2008 The stained nuclei can be analyzed by flow cytometry to monitor the level of gamma-H2AX to determine the level of DSBs and DNA content and A gamma-H2AX kit and antibody allows the assessment of DNA damage and DNA repair for ELISA based assays, immunohistochemistry or flow cytometry. X Phosphorylation Assay Kit (Flow cytometry) is a cell based assay formatted for flow cytometric detection of levels of phosphorylated Histone H2A.X.; find a signal for triggering DNA repair system so the γ-H2AX foci assay has potential use in flow cytometry is the indirect evidence of existence of DSBs. 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.

X, γ-H2AX, γH2AX Intracellular flow cytometry (3) applications for H2AX pS139 Antibody, anti-human/mouse, REAfinity™. Validated in WB, IHC, ICC, Flow Cyt and tested in Human. (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma; Flow Cytometry - Anti-gamma H2A. 26 Apr 2010 flow cytometry on a FACScan station with CellQuest software using the FL1 for FITC labeled caspase-3 active form or γ-H2AX. Western blotting. 20 Jun 2019 stress, both immunoblotting and flow cytometry analyses The γ-H2AX foci numbers per cell from 20-60 cells in each repeat of 3 biological 63 products gamma H2AX [p Ser139] Antibody · Applications: WB, FCM, ICC, IF, IHC, IHC-fr, IHC-p, ChIP · Reactivity: Hu, Ms, Rt, Ca · Conjugate/Tag: Unconjugated. P-H2AX flow cytometry assay in the clinic in a controlled manner.