γ h2ax detection kit  (Beyotime)

Journal: Toxics

Article Title: Oxidative Stress, DNA Damage, DNA Repair Inhibition, and Apoptosis Induced by Lead and Cadmium Combined Exposure in TK6 Cells

doi: 10.3390/toxics14040341

Figure Lengend Snippet: DNA damage markers induced by Pb and/or Cd in TK6 cells. ( A , C ) representative immunofluorescence staining images and quantification of γ-H2AX fluorescence intensity in TK6 cells. ( B , D , E ) representative the Comet assay images and quantitative analyses of Tail Length and Tail DNA%. Note: * p < 0.05, ** p < 0.01 for Pb and/or Cd group vs. the control group; while ## p < 0.01 for the single exposure groups vs. the mixed group; § p < 0.05, §§ p < 0.01 for the resveratrol group vs. the mixed group.

Article Snippet: DNA damage was further assessed using a γ-H2AX detection kit (Beyotime, Shanghai, China; C2035S), which quantifies γ-H2AX levels, a well-established biomarker for DNA double-strand breaks.

Techniques: Immunofluorescence, Staining, Fluorescence, Single Cell Gel Electrophoresis, Control

Journal: Scientific Reports

Article Title: ALDH18A1 has carcinogenic functions and regulates alternative splicing events of DNA repair-related genes in esophageal carcinoma cells

doi: 10.1038/s41598-025-08006-1

Figure Lengend Snippet: The si-ALDH18A1 group promotes DNA damage compared with the si-NC group in KYSE150 cells. ( A )The immunofluorescence imaging results of the DNA damage marker protein γ-H2AX, The green signal indicates DNA damage markers.The blue signal is DAPI, staining the nuclei. The Merge image shows the overlay of both channels. ( B )The quantitative comparison results of average fluorescence intensity between the two groups.*** P -value < 0.001.

Article Snippet: Detection of γ-H2AX protein expression by immunofluorescence: Aspirate the culture medium, wash once with PBS, add the fixative(1 ml per well for a 6-well plate)and fix for 5–15 min.After aspiration, wash three times with the washing solution(each time for 3–5 min), keeping the samples moist.Add the immunostaining blocking solution(1 ml per well for a 6-well plate)and block at room temperature for 10–20 min.After aspiration, add rabbit monoclonal antibody against γ-H2AX(1 ml per well for a 6-well plate,50 μl per well for a 96-well plate), and incubate at room temperature for 1 h or overnight at 4°C.Aspirate the antibody and save it.Wash three times with the washing solution(each time for 5–10 min), add anti-rabbit 488(1 ml per well for a 6-well plate)and incubate at room temperature for 1 h.Aspirate and save it.Wash twice with the washing solution(each time for 5–10 min), add DAPI staining solution(1 ml per well for a 6-well plate)and stain at room temperature for 5 min.After aspiration, wash three times(each time for 3–5 min).For a 6-well plate, add anti-fade mounting medium, cover with a coverslip, and observe under a fluorescence microscope.For a 96-well plate, the washing solution can be retained for direct observation or used for imaging analysis with a high-content analyzer.γ-H2AX staining shows green fluorescence, while DAPI staining of the cell nucleus shows blue fluorescence.The DNA damage detection kit is from Beyotime Biotechnology(C2035S).

Techniques: Immunofluorescence, Imaging, Marker, Staining, Comparison, Fluorescence