Tunel fluorescence

Manufactured by Promega

Sourced in United States

The TUNEL fluorescence assay is a method used to detect and quantify apoptosis, or programmed cell death, in cells. It works by labeling the fragmented DNA that is characteristic of apoptotic cells with a fluorescent dye, allowing for the identification and analysis of these cells using fluorescence microscopy or flow cytometry.

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2 protocols using tunel fluorescence

Studying RLIP76 Protein Function

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Purified 2HF (~99% pure) and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide (MTT) were purchased from Sigma–Aldrich (St. Louis, MO). Antibodies for CD31, Ki67, cyclin B1, CDK4, Bcl2, survivin, Bax, pAkt (S⁴⁷³), cleaved poly-ADP ribose polymerase (PARP), vimentin, and E-cadherin were purchased from Cell Signaling Technologies (Danvers, MA). Antibody for RLIP76 was purchased from Santa Cruz Biotechnology (Columbus, OH). Anti-rabbit and anti-mouse HRP-conjugated secondary antibodies were purchased from Cell Signaling Technologies. TUNEL fluorescence and CellTiter-Glo were procured from Promega (Madison, WI). Matrigel was purchased from Corning Life Sciences (Tewksbury, MA). Avidin/biotin complex (ABC) detection kit was procured from Vector (Burlingame, CA). The universal Mycoplasma detection kit was procured from ATCC (Manassas, VA). AlamarBlue was purchased from Thermo Fisher Scientific, Rockford, IL. Cell invasion assay kit was purchased from Cell Biolabs, Inc (San Diego, CA). RLIP76 shRNA and GFP-RLIP76 plasmid DNAs along with their respective controls were a kind gift from Dr Lawerence E. Goldfiner (Temple University, Philadelphia, PA).

Singhal J., Chikara S., Horne D., Salgia R., Awasthi S, & Singhal S.S. (2018). 2′-Hydroxyflavanone inhibits in vitro and in vivo growth of breast cancer cells by targeting RLIP76. Molecular carcinogenesis, 57(12), 1751-1762.

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Antibody-based Protein Expression Analysis

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2HF (purity ∼99%), Horseradish peroxidase (HRP)-conjugated anti-mouse, and anti-rabbit secondary antibodies, and MTT were purchased from Sigma-Aldrich, St. Louis, MO, USA. The CD31, Ki67, ERα, cyclin B1, CDK4, BCL-2, survivin, BIM, BAX, pAkt (S⁴⁷³), pERK (T^202/204), pSTAT3 (Y⁷⁰⁵), cleaved poly-ADP ribose polymerase (PARP), pJAK2 (Y^1007/1008), vimentin, fibronectin, and E-cadherin antibodies were purchased from Santa Cruz Biotechnology (Columbus, OH, USA) and Cell Signaling Technologies (Danvers, MA, USA). ELISA kit for VEGF expression was procured from R & D Systems (Minneapolis, MN, USA). TUNEL fluorescence and CellTiter-Glo were procured from Promega (Madison, WI, USA). Matrigel was purchased from Corning Life Sciences (Tewksbury, MA, USA). Avidin/biotin complex (ABC) detection kit was procured from Vector (Burlingame, CA, USA). The universal Mycoplasma detection kit was procured from ATCC (Manassas, VA, USA).

Singhal J., Nagaprashantha L., Chikara S., Awasthi S., Horne D, & Singhal S.S. (2017). 2'-Hydroxyflavanone: A novel strategy for targeting breast cancer. Oncotarget, 8(43), 75025-75037.

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