nb100-79967  (Bio-Techne corporation)

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

Journal: Frontiers in Public Health

Article Title: The Use of Genotoxicity Endpoints as Biomarkers of Low Dose Radiation Exposure in Interventional Cardiology

doi: 10.3389/fpubh.2021.701878

Figure Lengend Snippet: Mean values of chromosomal aberrations, frequency of Micronuclei (MN), and γ-H2AX foci per cell before the exposure, immediately after the interventional procedure ( in vivo radiation), and 24 h later, as well as the percentage of γ-H2AX foci repaired at 24 h.

Article Snippet: The main steps were the permeabilization of the cells, blocking of non-specific binding, immunostaining with primary γ-H2AX antibody (rabbit 1:1,000, Cat: NB100-79967, Novus Biologicals, Abingdon, UK) and secondary fluorescent antibody (Rhodamine Red-X anti-rabbit, 1:4,000, Cat: R6394, Life Technologies).

Techniques: In Vivo

Journal: Frontiers in Public Health

Article Title: The Use of Genotoxicity Endpoints as Biomarkers of Low Dose Radiation Exposure in Interventional Cardiology

doi: 10.3389/fpubh.2021.701878

Figure Lengend Snippet: Mean values of γ-H2AX foci and frequency of Micronuclei (MN) per cell after cardiac interventional procedures ( in vivo radiation) and after irradiation with 1 Gy in the laboratory ( in vitro radiation).

Article Snippet: The main steps were the permeabilization of the cells, blocking of non-specific binding, immunostaining with primary γ-H2AX antibody (rabbit 1:1,000, Cat: NB100-79967, Novus Biologicals, Abingdon, UK) and secondary fluorescent antibody (Rhodamine Red-X anti-rabbit, 1:4,000, Cat: R6394, Life Technologies).

Techniques: In Vivo, Irradiation, In Vitro

Journal: Andrology

Article Title: Spontaneous testicular atrophy occurs despite normal spermatogonial proliferation in a Tp53 knockout rat

doi: 10.1111/andr.12409

Figure Lengend Snippet: γH2AX levels do not differ between p53+/+ and p53−/− rat testes. Western blot analysis indicated no significant difference in γH2AX protein quantity (mean ± SEM) between p53+/+ and p53−/− rats (t-test, n = 5 rats) (A). Lanes labeled with “a” are at least 50% atrophic. Red boxes correspond with atrophic samples labeled on Western blot. Immunofluorescent images of testes stained for γH2AX (red) and Hoechst counterstain (blue) indicate that the pattern of γH2AX localization is similar in the wild-type testis (B, C, F–H) and non-atrophic p53-knockout testis tubules (D, E, I–K). In stages I–VI and VII–VIII (B–E), γH2AX is present in the sex vesicles of pachytene spermatocytes. During stages IX–XIV (F–K), a large number of γH2AX foci are present in leptotene spermatocytes (IX–XI), becoming more focused in zygotene spermatocytes (XII–XIII), and condensing into the punctate sex vesicle stain in early pachytene spermatocytes (XIV). Spermatogonia do not show evidence of γH2AX presence in either genotype (e.g. white arrowhead in D). Scale bar = 20 μm.

Article Snippet: Sections were incubated with monoclonal rabbit-anti-γH2AX antibody (Novus Biologicals, #NB100-79967), diluted 1:500 in blocking buffer, or blocking buffer alone as negative control, overnight at 4°C.

Techniques: Western Blot, Labeling, Staining, Knock-Out