rabbit polyclonal anti phospho gamma h2ax  (Novus Biologicals)

Journal: Journal of Medicinal Chemistry

Article Title: RTx-303, an Orally Bioavailable Polθ Polymerase Inhibitor That Potentiates PARP Inhibitors in BRCA Mutant Tumors

doi: 10.1021/acs.jmedchem.5c00551

Figure Lengend Snippet: Cellular and in vitro activity of RTx-284. A. Bar plot showing the IC50 of RTx-284 in the indicated HR-proficient (gray) and HRD (red) cell lines. Data represent mean of at least 3 independent experiments performed in triplicate. B-D. Synergy plots created by Combenefit showing synergistic activity between RTx-284 and indicated PARPi in the indicated HRD cell lines. E. Bar plot showing that 10 μM RTx-284 suppresses the MMEJ GFP reporter. Data represent mean of 3 individual experiments performed in triplicate ±SEM. F. Dot plot showing increase of γH2AX following RTx-284:olaparib treatment in DLD1 BRCA2–/– cells. G. Bar plot showing that RTx-284 promotes an increase in caspase positive DLD1 BRCA2–/– cells. Data represent mean of 3 independent experiments performed in triplicate, ±SEM.

Article Snippet: Cells were incubated with primary antibody (rabbit antigamma H2AX (p Ser139) antibody, Bethyl Lab #A700–053, 1:500 dilution in 1% BSA in PBS) overnight at 4 °C followed by 3x washes with PBS and then 1 h incubation with secondary antibody (Goat anti-Rabbit IgG (H+L) Secondary Antibody, DyLight 488 (Thermo #35552) 1:2000 dilution in 1% BSA in PBS).

Techniques: In Vitro, Activity Assay

Journal: Scientific Reports

Article Title: Acute ischemic stroke triggers a cellular senescence-associated secretory phenotype

doi: 10.1038/s41598-021-95344-5

Figure Lengend Snippet: Microglial cells of the ischemic area showed a higher expression of NF-kB and γ-H2AX. ( A ) Immunofluorescence NF-kB expression showed an infarct area location, delimited by white dotted line. Plot represented the NF-kB expression by areas (contralateral vs infarct area: p < 0.0001; ipsilateral vs infarct: p = 0.0009; contralateral vs ipsilateral: p < 0.0001). Scale bar = 100 µm; insert scale bar = 40 µm. ( B ) Nuclear expression of γ-H2AX in microglial cells. Plot represented the γ-H2AX positive foci in contralateral, ipsilateral and infarct areas (contralateral vs infarct: p < 0.0001; ipsilateral vs infarct: p = 0.0004). Scale bar = 100 µm; insert scale bar = 50 µm.

Article Snippet: Immunohistochemistry was performed against mouse anti-p16 (1:200; Abcam, Cat. No. ab54210), mouse anti-p21 (1:200; Santa Cruz Biotechnology, Cat. No. sc-817), rabbit anti-GFAP (1:200; Abcam, Cat. No. ab7260), rabbit anti-Iba1 (1:200; FUJIFILM Wako, Cat. No. 019-19741), rabbit anti-NeuN (1:200; Abcam, Cat.No.ab177487), rabbit anti-NF-κB p65 (1:200; Proteintech, Cat. No. 10745-1-AP), chicken anti-Iba1 (1:200; Synaptic Systems, Cat No. 234 006), and rabbit gamma H2AX [p Ser139] antibody (EP854(2)Y, 1:200; NOVUS Biologicals, Cat. No. NB100-79967) primary antibodies and followed by appropriated secondary antibodies.

Techniques: Expressing, Immunofluorescence