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Cell Meter 通用Caspase活性熒光法檢測試劑盒 紅色熒光 適合于流式細胞檢測

貨號22822存儲條件 在零下15度以下保存, 避免光照
規格100 Tests價格2544
Ex (nm)649Em (nm)664
分子量溶劑
產品詳細介紹


簡要概述

我們的Cell Meter 檢測試劑盒是一套用于檢測細胞生存力的工具。有多種參數可用于檢測細胞活力。該特定試劑盒旨在通過測量活細胞中的通用胱天蛋白酶(胱天蛋白酶-1,-3,-4,-5,-6,-7,-8和-9)激活來檢測細胞凋亡。胱天蛋白酶激活被廣泛認為是細胞凋亡的可靠指標。大多數胱天蛋白酶具有對肽序列Val-Ala-Asp(VAD)的底物選擇性。該試劑盒使用TF5-VAD-FMK作為大多數半胱天冬酶活性的熒光指示劑。 TF5-VAD-FMK是細胞可滲透的,無毒的,并且與凋亡細胞中活化的casepase-1,-3,-4,-5,-6,-7,-8和-9不可逆地結合。一旦與胱天蛋白酶結合,紅色熒光試劑將保留在細胞內。結合后阻止了胱天蛋白酶的進一步催化,但不會阻止凋亡的進行。添加到培養基中后的15分鐘內,該試劑將開始與活性caspase酶反應。該試劑盒為所有必需成分提供了優化的測定方案。它用于定量凋亡細胞中大多數激活的胱天蛋白酶活性,或篩選胱天蛋白酶抑制劑。紅色熒光標記允許通過流式細胞儀直接檢測凋亡細胞中激活的胱天蛋白酶。百螢生物是AAT Bioquest的中國代理商,為您提供最優質的Cell Meter 通用Caspase活性熒光法檢測試劑盒。

點擊查看光譜

 

適用儀器


流式細胞儀  
激發: 488nm激發
發射: 530/30nm濾波片
通道: FITC通道


產品說明書

樣品實驗方案

簡要概述

  1. 用5×105至1×106細胞/ mL的密度制備含測試化合物的細胞
  2. 將0.5 µL細胞溶液中加入1 µL 500X TF5-VAD-FMK
  3. 將細胞在37°C,5%CO2培養箱中孵育1-4小時
  4. 沉淀細胞并將細胞重懸于0.5 mL檢測緩沖液或生長培養基中
  5. 使用帶有660/20 nm濾光片的流式細胞儀分析細胞(APC通道)

 

實驗步驟

1.對于每個樣品,在0.5mL溫熱培養基或自備緩沖液中以5×105至1×106cell/ mL的密度制備細胞。注意:應單獨評估每種細胞系,以確定誘導凋亡的最佳細胞密度。

2.用測試化合物處理細胞一段時間,以誘導細胞凋亡,并建立陽性和陰性對照。

3.向處理過的細胞中加入1µL 500X TF5-VAD-FMK(組分A)。

4.將細胞在37°C,5%CO2培養箱中孵育1-4小時。注意:對于貼壁細胞,用0.5 mM EDTA輕輕提起細胞以保持細胞完整,并在與TF5-VAD-FMK孵育之前用含血清的培養基洗滌細胞一次。合適的孵育時間取決于所用的單個細胞類型和細胞濃度。優化每個實驗的孵育時間。

5.洗滌并旋轉細胞兩次。將細胞重懸于0.5 mL分析緩沖液(組分B)或生長培養基中。注意:TF5-VAD-FMK是熒光的;因此,重要的是洗凈任何未結合的試劑以除去背景。

6.可選:用DNA染色劑標記細胞(例如綠色DCS標記死細胞,可以用530/30 nm濾光片(FITC通道)檢測到。

7.可選:修復細胞。

8.使用帶有660/20 nm濾光片(APC通道)的流式細胞儀檢測熒光強度。

 

參考文獻

Angiopoietins Modulate Survival, Migration, and the Components of the Ang-Tie2 Pathway of Chronic Lymphocytic Leukaemia (CLL) Cells In Vitro
Authors: Palma, Luis Mario Aguirre and Flamme, Hanna and Gerke, Iris and Kreuzer, Karl-Anton
Journal: Cancer Microenvironment (2016): 13--26

Hypoxia regulates TRAIL sensitivity of colorectal cancer cells through mitochondrial autophagy.
Authors: Knoll, Gertrud and Bittner, Sebastian and Kurz, Maria and Jantsch, Jonathan and Ehrenschwender, Martin
Journal: Oncotarget (2016)

Promotion of osteointegration under diabetic conditions by tantalum coating-based surface modification on 3-dimensional printed porous titanium implants
Authors: Wang, Lin and Hu, Xiaofan and Ma, Xiangyu and Ma, Zhensheng and Zhang, Yang and Lu, Yizhao and Li, Xiang and Lei, Wei and Feng, Yafei
Journal: Colloids and Surfaces B: Biointerfaces (2016): 440--452

Role of delta-like ligand-4 in chemoresistance against docetaxel in MCF-7 cells
Authors: Wang, Q and Shi, Y and Butler, HJ and Xue, J and Wang, G and Duan, P and Zheng, H
Journal: Human & Experimental Toxicology (2016): 0960327116650006

microRNA-186 inhibits cell proliferation and induces apoptosis in human esophageal squamous cell carcinoma by targeting SKP2
Authors: He, Wei and Feng, Jianfang and Zhang, Yan and Wang, Yuanyuan and Zang, Wenqiao and Zhao, Guoqiang
Journal: Laboratory Investigation (2016): 317--324

Glucose promotes cell proliferation, glucose uptake and invasion in endometrial cancer cells via AMPK/mTOR/S6 and MAPK signaling
Authors: Han, Jianjun and Zhang, Lu and Guo, Hui and Wysham, Weiya Z and Roque, Dario R and Willson, Adam K and Sheng, Xiugui and Zhou, Chunxiao and Bae-Jump, Victoria L
Journal: Gynecologic oncology (2015): 668--675

IL-7 abrogates the immunosuppressive function of human double-negative T cells by activating Akt/mTOR signaling
Authors: Allgäuer, Andrea and Schreiner, Elisabeth and Ferrazzi, Fulvia and Ekici, Arif B and Gerbitz, Armin and Mackensen, Andreas and Völkl, Simon
Journal: The Journal of Immunology (2015): 3139--3148

Insulin improves osteogenesis of titanium implants under diabetic conditions by inhibiting reactive oxygen species overproduction via the PI3K-Akt pathway
Authors: Wang, Lin and Zhao, Xiong and Wei, Bo-yuan and Liu, Yi and Ma, Xiang-yu and Wang, Jian and Cao, Peng-chong and Zhang, Yang and Yan, Ya-bo and Lei, Wei and others
Journal: Biochimie (2015): 85--93

LGL1 modulates proliferation, apoptosis, and migration of human fetal lung fibroblasts
Authors: Zhang, Hui and Sweezey, Neil B and Kaplan, Feige
Journal: American Journal of Physiology-Lung Cellular and Molecular Physiology (2015): L391--L402

t-BHQ Provides Protection against Lead Neurotoxicity via Nrf2/HO-1 Pathway
Authors: Ye, Fang and Li, Xiaoyi and Li, Lili and Yuan, Jing and Chen, Jun
Journal: Oxidative medicine and cellular longevity (2015)



說明書
Cell Meter 通用Caspase活性熒光法檢測試劑盒 紅色熒光 適合于流式細胞檢測 .pdf


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