γ h2ax

Manufactured by Abways

Sourced in China, United States

The γ-H2AX is a lab equipment product used to detect DNA double-strand breaks. It functions by measuring the phosphorylation of the histone protein H2AX, which is a marker for the presence of DNA damage.

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Lab products found in correlation

H3k27me3, by Merck Group (1 mentions) Gsk j4, by Selleck Chemicals (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Hct116, by American Type Culture Collection (1 mentions) Sw620, by American Type Culture Collection (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by R&D Systems (1 mentions) Oxaliplatin, by Selleck Chemicals (1 mentions) H3k4me3, by Active Motif (1 mentions) Epz 6438, by Selleck Chemicals (1 mentions) H3k27me3, by Cell Signaling Technology (1 mentions) Cleaved caspase3, by Abways (1 mentions) P bcl 2, by Cell Signaling Technology (1 mentions) P erk, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Bcl 2, by Cell Signaling Technology (1 mentions) H3k27me3, by Active Motif (1 mentions) Triton x 100, by Biosharp (1 mentions) Goat anti rabbit igg h l alexa fluor 488, by Abways (1 mentions) Nlrp3, by Abways (1 mentions) Caspase 1 p20, by Affinity Biosciences (1 mentions)

4 protocols using γ h2ax

Colorectal Cancer Cell Line Profiling

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Human CRC cell lines HCT116 and SW620 were purchased from American Type Culture Collection. Cells were cultured in Dulbecco’s modified Eagle medium containing 10% fetal calf serum medium. All CRC cells were negative for mycoplasma contamination prior to use.
Oxaliplatin, GSK-J4, and EPZ-6438 were purchased from Selleck. DAPI (#5748) was obtained from R&D Systems. The following antibodies were used: H3K27me3 (9733, Cell Signaling Technology) for IHC, H3K27me3 (07–449; Millipore) for ChIP and western blot, H3K4me3 (61379, ActiveMotif), H3 (sc-10809, Santa Cruz), PAPR (9542, Cell Signaling Technology), γH2A.X (3322, Abways), and cleaved caspase 3 (9664, Cell Signaling Technology).

Wang Q., Chen X., Jiang Y., Liu S., Liu H., Sun X., Zhang H., Liu Z., Tao Y., Li C., Hu Y., Liu D., Ye D., Liu Y., Wang M, & Zhang X. (2019). Elevating H3K27me3 level sensitizes colorectal cancer to oxaliplatin. Journal of Molecular Cell Biology, 12(2), 125-137.

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Western Blot Analysis of Cellular Proteins

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Cells were dissolved in lysis buffer containing protease inhibitors. Then, the proteins were subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to 0.22 μm polyvinylidene fluoride (PVDF) membranes. The membranes were blocked with 5% nonfat milk at room temperature for 2 h and then incubated with primary antibodies, including antibodies against GAPDH (Cell Signaling Technology), H3K27me3 (Active Motif), H3 (Active Motif), PRKCA (Abways), BCL2 (Cell Signaling Technology), pBCL2 (Cell Signaling Technology), ERK (Cell Signaling Technology), pERK (Cell Signaling Technology), RAF (Abways), pRAF (Abways), γH2AX (Abways), SOX9 (Abways), CD117 (Abways), and cleaved Caspase3 (Abways), overnight at 4 °C. After three washes in Tris-buffered saline with Tween 20 (TBST), the membranes were incubated with corresponding secondary antibodies for 2 h at room temperature.

He C., Sun J., Liu C., Jiang Y, & Hao Y. (2019). Elevated H3K27me3 levels sensitize osteosarcoma to cisplatin. Clinical Epigenetics, 11, 8.

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Immunofluorescence Analysis of Aortic NLRP3 Inflammasome

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Mice thoracic aortas were separated, optimal cutting temperature compound (OCT, CellPath, U.K.) embedded aorta frozen sections were prepared, and immunofluorescence staining was performed as described previously [23] . Frozen sections were fixed by 4% paraformaldehyde for 15 min, then permeabilized by 0.3% (v/v) Triton X-100 (Biosharp, Hefei, China) in PBS for 15 min and blocked with 5% (w/v) bovine serum albumin (BSA, Solarbio, Beijing, China) for 1 h, then incubated with primary antibodies against NLRP3 (1:100, Abways Technology, Inc., Shanghai, China), PYCARD (ASC, 1:100, Abways Technology, Inc., Shanghai, China), Caspase-1 p20 (1:100, Affinity Biosciences, OH, USA) and tert (1:100, Affinity Biosciences, OH, USA) at 4°C overnight, followed by Cy3-conjugated Goat Anti-Rabbit IgG (H + L) (1:100, Proteintech™, Wuhan, China) and Goat Anti-Rabbit IgG (H + L) Alexa Fluor 488 (1:100, Abways Technology, Inc., Shanghai, China) for 2 h, respectively. DAPI was used to stain the nucleus.
Treated VSMCs and ECs were fixed and blocked like the above procedures, and then incubated with γ-H2AX (1:100, Abways Technology, Inc., Shanghai, China), followed by Cy3, and then counterstained with DAPI.

Tai G.J., Yu Q.Q., Li J.P., Wei W., Ji X.M., Zheng R.F., Li X.X., Wei L, & Xu M. (2022). NLRP3 inflammasome links vascular senescence to diabetic vascular lesions. Pharmacological research, 178.

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Immunohistochemical Analysis of Cellular Markers

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The fixed thoracic aorta was embedded with paraffin and 4-6 µm thick sections were prepared. After the slides were deparaffinized and hydrated, antigen retrieval was conducted, then permeabilized and the peroxidase was inactivated by 3% H 2 O 2 . Tissue was blocked by 5% goat serum (Beyotime Biotechnology, Shanghai, China) in PBS for 1 h and incubated with primary antibodies of p53 (1:200, Wanleibio, Shengyang, China), p21 Cip1 (1:200, Affinity Biosciences, OH, USA), γ-H2AX (1:100, Abways Technology, Inc., Shanghai, China) at 4 °C for 12 h, after washing, horseradish peroxidase-conjugated rabbit anti-goat IgG (1:500, Abways Technology, Inc., Shanghai, China) was applied for 30 min at room temperature. Finally, the staining was performed by using DAB kit (ZSGB-BIO, Beijing, China), followed by counterstaining with hematoxylin.

Tai G.J., Yu Q.Q., Li J.P., Wei W., Ji X.M., Zheng R.F., Li X.X., Wei L, & Xu M. (2022). NLRP3 inflammasome links vascular senescence to diabetic vascular lesions. Pharmacological research, 178.

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