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    SA-b-gal染色實驗

    SA-b-gal染色實驗檢測目的基因消減對細胞衰老的影響

    技術原理

    大多數正常細胞在經過有限次分裂后會進入衰老期,不能再在一些常規刺激的誘導下進行分裂。其細胞周期,基因和蛋白表達都發生了改變。衰老的細胞體積會變大,在pH6.0時會表達高酶活性的β-半乳糖苷酶。利用細胞衰老時產生的β-半乳糖苷酶催化底物后,在光學顯微鏡下對細胞進行拍照計數,從而反映處于衰老狀態的細胞數量。

    結果展示


    通過SA-b-gal染色實驗檢測siRNA慢病毒感染后,實驗組促進衰老樣表型發生。

    參考文獻

    1.Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O (September 1995). "A biomarker that identifies senescent human cells inculture and in aging skin in vivo". Proc. Natl. Acad. Sci. U.S.A.92 (20): 9363–7.

    2.Bassaneze V, Miyakawa AA, Krieger JE (January 2008). "A quantitative chemiluminescent method for studying replicative and stress-induced premature senescence in cell cultures". Anal. Biochem.372 (2): 198–203.

    3.Gary RK, Kindell SM (August 2005). "Quantitative assay of senescence-associated beta-galactosidase activity in mammalian cell extracts". Anal. Biochem.343 (2): 329–34.

    4.Itahana K, Campisi J, Dimri GP (2007). "Methods to detect biomarkers of cellular senescence: the senescence-associated beta-galactosidase assay". Methods Mol. Biol. Methods in Molecular Biology 371: 21–31.doi:10.1007/978-1-59745-361-5_3.

    5.Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D, Hwang ES (April 2006). "Senescence-associated beta-galactosidase is lysosomal beta-galactosidase". Aging Cell5 (2): 187–95.

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