Γh2ax Staining
Γh2ax Staining - Related Videos
04:09
Assessing DNA Damage Foci: To Identify Radiation-Induced DNA Damage Foci in Cancer Cell Line Using Immunofluorescence Staining
0 Views •
2023
This video describes a protocol to detect radiation-induced DNA repair foci in the colon cancer cell lines using immunofluorescence staining. DNA repair proteins get activated at the DNA damage site; therefore, we can detect them using specific antibodies and visualize them under a fluorescence microscope.
11:33
Microscopy and Staining: Gram, Capsule, and Endospore Staining
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Source: Rhiannon M. LeVeque1, Natalia Martin1, Andrew J. Van Alst1, and Victor J. DiRita1 1 Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, United States of America Bacteria are diverse microorganisms found nearly everywhere on Earth. Many properties help distinguish them from each other, including but not limited to Gram-staining type, shape and arrangement, production of capsule, and formation of spores. To observe these properties, one...
03:12
May-Grunwald Giemsa Staining: A Method to Stain Bone Marrow Cells
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10:55
Staining Proteins in Gels
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Cited by 7 •
2008
Following separation by electrophoretic methods, proteins in a gel can be detected by several staining methods. Staining of proteins with Coomassie Blue, Silver Staining, SYPRO Orange, SYPRO Ruby are demonstrated in this video.
08:23
An Automated Microscopic Scoring Method for the γ-H2AX Foci Assay in Human Peripheral Blood Lymphocytes
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Cited by 7 •
2021
This protocol presents the slide preparation and automated scoring of the γ-H2AX foci assay on peripheral blood lymphocytes. To illustrate the method and sensitivity of the assay, isolated lymphocytes were irradiated in vitro. This automated method of DNA DSB detection is useful for fast and high-throughput biological dosimetry applications.
FAQs
The signal begins when DNA damage activates phosphatidylinositol 3-kinase-related kinases, which phosphorylate histone H2AX at serine 139. This modified form, γH2AX, accumulates around the affected chromatin, creating a localized molecular mark that antibodies can detect. The phosphorylation step connects damage-associated signaling with the visible staining pattern used for analysis.
Nuclear foci provide a spatial readout rather than only a bulk indication of cellular stress. Because γH2AX accumulates around damaged chromatin, researchers can examine these localized signals in individual nuclei or tissue regions. Measuring the foci supports comparisons of DNA damage among experimental conditions, including cells exposed to radiation or chemotherapy.
Patterns of γH2AX foci can help compare how tumor cells respond to DNA-damaging treatments and how their repair responses differ. The method does not simply record that treatment occurred; it provides a staining-based damage readout that can be examined across conditions. In cancer research, this supports evaluation of treatment-associated genomic damage and response patterns.
The two antibody-based formats support related γH2AX measurements but suit different sample contexts. Immunofluorescence is used to visualize foci in cells, whereas immunohistochemistry supports analysis of tissue samples. Selecting between them allows investigators to match the detection approach to the material being studied while retaining the same damage-associated marker.
A basic γH2AX staining workflow begins with a cell or tissue sample and uses antibody-based immunofluorescence or immunohistochemistry to detect the phosphorylated marker. Researchers then examine or measure the resulting nuclear foci. Applying the same readout across experimental conditions allows comparisons of treatment-associated DNA damage, repair responses, or genotoxic effects.
In cancer research, γH2AX staining can assess how tumor cells respond to radiation or chemotherapy and compare their repair responses across experimental conditions. It also supports investigation of genotoxic effects, meaning damage to genetic material, and treatment-associated genomic damage. Together, these uses connect a molecular staining readout with cancer-treatment research questions.