中英文說明書丨艾美捷細(xì)胞衰老β-半乳糖苷酶染色試劑盒
The Cellular Senescence Assay Kit provides an easy-to-use and efficient method to determine cellular
senescence. SA-β-galactosidase catalyzes the hydrolysis of X-gal, which produces a blue color. Each
kit provides sufficient quantities to perform up to 50 assays in 35 mm dishes.
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Kit Components
1. 100X Fixing Solution (Part No. 40010): One tube – 1.5 mL of 25% Glutaraldehyde
2. Staining Solution A (Part No. 40011): One tube – 1.5 mL of 500 mM Potassium Ferrocyanide
3. Staining Solution B (Part No. 40012): One tube – 1.5 mL of 500 mM Potassium Ferricyanide
4. Staining Solution C (Part No. 40015): One bottle – 4.5 mL of 1 M Citrate-Na2HPO4 Buffer, pH
6.0, 50 mM MgCl2
5. Staining Solution D (Part No. 40016): One bottle – 4.0 mL of 5 M NaCl
6. X-gal Solution (Part No. 40014): Two tubes – 1.5 mL of 40 mg/mL X-gal in DMF in each tube
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Storage
Store X-gal solution protected from light at -20oC. Store all other components at 4oC.
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Preparation of Reagents
? 1X Fixing Solution: Prepare a 1X Fixing Solution by diluting the provided 100X stock 1:100 in 1X
PBS. Store the diluted solution at room temperature for up to six months.
? Cell Staining Working Solution: Prepare FRESH cell staining working solution based on the
number of samples. The chart below is suggested for samples in 35 mm plate, and may be
modified accordingly to suit other culture plate sizes.
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艾美捷細(xì)胞衰老β-半乳糖苷酶染色試劑盒提供了一種有效的方法來可視化衰老相關(guān)（SA）?-半乳糖苷酶。SA-?-Gal催化X-gal的水解，在衰老細(xì)胞中產(chǎn)生藍(lán)色。使用標(biāo)準(zhǔn)光學(xué)顯微鏡可視化結(jié)果。
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艾美捷細(xì)胞衰老β-半乳糖苷酶染色試劑盒基本參數(shù)：
全稱：細(xì)胞衰老檢測試劑盒（細(xì)胞衰老相關(guān)β-半乳糖苷酶染色法）
英文：Cellular Senescence Detection Kit (SA-β-Gal Staining)
編號：CBA-230
規(guī)格：50assays/5 x 50 assays
保存：4oC
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艾美捷細(xì)胞衰老β-半乳糖苷酶染色試劑盒試劑的制備
?1X固定溶液：通過將提供的100X原液稀釋成1:100的1X溶液，制備1X固定溶液
PBS。將稀釋的溶液在室溫下儲存六個月。
?細(xì)胞染色工作溶液：根據(jù)
樣本數(shù)量。以下圖表適用于35mm板中的樣品
相應(yīng)地修改以適合其他培養(yǎng)板尺寸。
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分析方案（35mm皿）：
1.從表達(dá)SA-?-Gal的衰老細(xì)胞中吸出培養(yǎng)基。
2.用3 mL 1X PBS洗滌細(xì)胞兩次，并吸出最終洗滌液。
3.加入2mL 1X固定溶液。在室溫下孵育5分鐘。
4.取出固定溶液，用3mL 1X PBS洗滌固定細(xì)胞三次。
5.吸入最后一次洗滌液，并通過添加2 mL新鮮制備的細(xì)胞完全覆蓋細(xì)胞染色工作溶液。
6.在37oC避光條件下培養(yǎng)細(xì)胞4小時至過夜。
7.取出細(xì)胞染色工作溶液，然后用3 mL 1X洗滌染色細(xì)胞兩次PBS并將細(xì)胞儲存在1X PBS中。對于長期儲存，用1X PBS覆蓋細(xì)胞20%甘油。儲存在4oC。
注：通過將染色樣品與DMSO。
8.?使用光學(xué)顯微鏡計數(shù)藍(lán)色染色的衰老細(xì)胞。
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細(xì)胞衰老β-半乳糖苷酶染色試劑盒文獻(xiàn)參考：
1. Current Protocols in Molecular Biology, John Wiley & Sons Press.
2. Campisi, J. (2000) In Vivo 14, 183-188.
3. Dimri, G. P., X. Lee, G. Basile, M. Acosta, G. Scott, C. Roskelley, E. E. Medrano, M. Linskens, I.
Rubelj, O. Pereira-Smith, M. Peacocke, and J. Campisi. (1995) Proc. Natl. Acad. Sci. USA
92:9363-9367.
