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Cell Meter TUNEL凋亡檢測試劑盒 綠色熒光

貨號22849存儲條件 在零下15度以下保存, 避免光照
規格50 Tests價格3840
Ex (nm)498Em (nm)522
分子量溶劑
產品詳細介紹


簡要概述

DNA片段化代表晚期細胞凋亡的特征。凋亡細胞中的DNA斷裂可以通過末端脫氧核苷酸轉移酶(TdT)介導的dUTP缺口末端標記(TUNEL)檢測。 TUNEL測定法依賴于DNA中存在的缺口,該缺口可通過TdT進行鑒定,TdT是一種酶,該酶催化添加標記物的dUTP?,F有的所有TUNEL分析均包含劇毒的甲胂酸鈉,這可能會誘導細胞凋亡并降低DNA的產生和DNA鏈。我們的Cell Meter TUNEL細胞凋亡測定試劑盒使用了不含甲胂酸鈉的專有緩沖系統。該試劑盒基于將我們專有的熒光染料摻入凋亡過程中形成的DNA片段中。該測定法經過優化,可直接檢測分離的或貼壁細胞中的細胞凋亡,而無需使用任何抗體。該試劑盒為所有必需成分提供了優化的測定方案。適用于熒光酶標儀,熒光顯微鏡或流式細胞儀。在流行的FITC通道上可以輕松檢測到其信號。百螢生物是AAT Bioquest的中國代理商,為您提供最優質的Cell Meter TUNEL凋亡檢測試劑盒。

點擊查看光譜

 

適用儀器


流式細胞儀  
激發: 488nm激光
發射: 530/30nm濾波片
通道: FITC濾波片
熒光顯微鏡  
激發: FITC濾波片
發射: FITC濾波片
推薦孔板: 黑色透明
熒光酶標儀  
激發: 490nm
發射: 525nm
cutoff: 515nm
推薦孔板: 黑色孔板


產品說明書

樣品實驗方案

簡要概述

  1. 用測試化合物準備細胞。
  2. 與TUNEL工作溶液在37°C下孵育30分鐘至1小時。
  3. 洗滌細胞。
  4. 用4%甲醛(可選)固定細胞。
  5. 用帶FITC濾光片的熒光顯微鏡或帶FITC通道的流式細胞儀。讀取Ex / Em = 490/525 nm(截止= 515 nm)處的熒光強度。

 

溶液配制 

工作溶液配制

將0.5μL的100X Tunnelyte Green(組分A)添加到50μL的反應緩沖液(組分B)中,使總體積為50.5μL的TUNEL工作溶液。 避光。 注意:應單獨評估每個細胞系,以確定最佳細胞密度。

 

實驗步驟

1.根據您的特定協議,將細胞培養至最佳密度以誘導凋亡。 對于在96孔板培養物中生長的貼壁細胞,我們建議大約30,000至50,000個細胞/孔,對于非貼壁細胞,建議大約1至2 x 106細胞/ mL。 同時,在每種標記條件下,用誘導群體相同的密度培養非誘導陰性對照細胞群體。 注意:我們用100 nM-1 µM星形孢菌素處理HeLa細胞4小時,以誘導細胞凋亡。

2.染色和固色:

2.1取出細胞培養基。
2.2向每個樣品中添加50µL TUNEL工作溶液。
2.3在37°C下孵育30-60分鐘。
2.4除去TUNEL工作溶液,并用200 µL /孔的PBS洗滌細胞1-2次。
2.5向每個樣品中添加100uL反應緩沖液(組分B)。
2.6使用Ex / Em = 490/525 nm(cut off= 515 nm)的熒光酶標儀,帶有FITC濾光片組的熒光顯微鏡或帶有FITC通道的流式細胞儀檢測熒光強度。
2.7可選:從步驟5中移出反應緩沖液,并向每個孔中添加100 µL /孔/ 96孔板的4%甲醛固定緩沖液(未提供)。注意:對于非貼壁細胞,請添加所需量(例如2X106細胞/ mL)的4%甲醛固定液。
2.8在室溫下將平板孵育20至30分鐘。
2.9去除固定劑。
2.10用PBS洗滌細胞2-3次,并用100µL PBS /孔/ 96孔板替換。
2.11使用Ex / Em = 490/525 nm(cut off= 515 nm)的熒光酶標儀,帶有FITC濾光片組的熒光顯微鏡或帶有FITC通道的流式細胞儀檢測熒光強度。
2.12可選:用1X Hoechst(組分C)在Ex / Em = 350/460 nm處染色細胞核,以進行圖像分析

 

參考文獻

Vaccarin alleviates hypertension and nephropathy in renovascular hypertensive rats
Authors: Cai, Weiwei and Zhang, Zhenpeng and Huang, Yiqi and Sun, Haijian and Qiu, Liying
Journal: Experimental and Therapeutic Medicine (2018): 924--932

CO-releasing molecules-2 attenuates ox-LDL-induced injury in HUVECs by ameliorating mitochondrial function and inhibiting Wnt/β-catenin pathway
Authors: Sun, Hai-Jian and Xu, Dong-Yan and Sun, Yi-Xin and Xue, Tong and Zhang, Chen-Xing and Zhang, Zhi-Xuan and Lin, Wei and Li, Ke-Xue
Journal: Biochemical and Biophysical Research Communications (2017)

Salusin-β mediates high glucose-induced endothelial injury via disruption of AMPK signaling pathway
Authors: Zhu, Xuexue and Zhou, Yuetao and Cai, Weiwei and Sun, Haijian and Qiu, Liying
Journal: Biochemical and Biophysical Research Communications (2017)

Vaccarin protects human microvascular endothelial cells from apoptosis via attenuation of HDAC1 and oxidative stress
Authors: Zhu, Xuexue and Lei, Yueyue and Tan, Fanggen and Gong, Leilei and Gong, Haifeng and Yang, Wei and Chen, Ting and Zhang, Zhixuan and Cai, Weiwei and Hou, Bao and others
Journal: European Journal of Pharmacology (2017)

Axl is required for TGF-β2-induced dormancy of prostate cancer cells in the bone marrow
Authors: Yumoto, Kenji and Eber, Matthew R and Wang, Jingcheng and Cackowski, Frank C and Decker, Ann M and Lee, Eunsohl and Nobre, Ana Rita and Aguirre-Ghiso, Julio A and Jung, Younghun and Taichman, Russell S
Journal: Scientific Reports (2016)

Growth Arrest-Specific 6 (GAS6) Promotes Prostate Cancer Survival by G1 Arrest/S Phase Delay and Inhibition of Apoptotic Pathway During Chemotherapy in Bone Marrow
Authors: Lee, Eunsohl and Decker, Ann M and Cackowski, Frank C and Kana, Lulia A and Yumoto, Kenji and Jung, Younghun and Wang, Jingcheng and Buttitta, Laura and Morgan, Todd M and Taichman, Russell S
Journal: Journal of cellular biochemistry (2016)

RFX1--dependent activation of SHP-1 induces autophagy by a novel obatoclax derivative in hepatocellular carcinoma cells
Authors: Su, Jung-Chen and Tseng, Ping-Hui and Hsu, Cheng-Yi and Tai, Wei-Tien and Huang, Jui-Wen and Ko, Ching-Huai and Lin, Mai-Wei and Liu, Chun-Yu and Chen, Kuen-Feng and Shiau, Chung-Wai
Journal: Oncotarget (2014): 4909

In situ detection of apoptosis by the TUNEL assay: an overview of techniques
Authors: Loo DT.
Journal: Methods Mol Biol (2011): 3

Testicular apoptosis after dietary zinc deficiency: ultrastructural and TUNEL studies
Authors: Kumari D, Nair N, Bedwal RS.
Journal: Syst Biol Reprod Med (2011): 233

In situ localization of apoptosis using TUNEL
Authors: Hewitson TD, Darby IA.
Journal: Methods Mol Biol (2010): 161



說明書
Cell Meter TUNEL凋亡檢測試劑盒 綠色熒光 .pdf


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