Cell Meter 通用Caspase活性荧光法检测试剂盒

 

适用仪器

样品实验方案

简要概述

1. 用5×10⁵至1×10⁶细胞/ mL的密度制备含测试化合物的细胞
2. 将0.5 µL细胞溶液中加入1 µL 500X TF2-VAD-FMK
3. 将细胞在37°C，5％CO₂培养箱中孵育1-4小时
4. 沉淀细胞并将细胞重悬于0.5 mL检测缓冲液或生长培养基中
5. 使用带有530/30 nm滤光片（FITC通道）的流式细胞仪分析细胞

 

实验步骤

1.对于每个样品，在0.5 mL温热培养基或自备缓冲液中以5×10⁵至1×10⁶cell/ mL的密度制备细胞。注意：应单独评估每种细胞系，以确定诱导凋亡的佳细胞密度。

2.用测试化合物处理细胞一段时间，以诱导细胞凋亡，并建立阳性和阴性对照。

3.向处理过的细胞中加入1µL 500X TF2-VAD-FMK（组分A）。

4.将细胞在37°C，5％CO₂培养箱中孵育1-4小时。注意：对于贴壁细胞，用0.5 mM EDTA轻轻提起细胞以保持细胞完整，并在与TF2-VAD-FMK孵育之前用含血清的培养基洗涤细胞一次。合适的孵育时间取决于所用的单个细胞类型和细胞浓度。优化每个实验的孵育时间。

5.洗涤并旋转细胞两次。将细胞重悬于0.5 mL分析缓冲液（组分B）或生长培养基中。注意：TF2-VAD-FMK是荧光的；因此，重要的是洗净任何未结合的试剂以除去背景。

6.可选：可以用DNA染色剂标记细胞（例如碘化丙啶或死细胞的7-AAD）。

7.可选：修复细胞。

8.使用带有530/30 nm滤光片的流式细胞仪（FITC通道）检测荧光强度。

 

相关产品

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)