CYTO-ID^® 自噬检测试剂盒

◆特点

　　●　无需转染，定量监测活细胞中的自噬情况

　　●　免去LC3-GFP转染所需的时间和精力以及转染效率的验证

　　●　含有特殊基团的专利染料选择性地染色自噬小泡

　　●　试剂盒中包含已知活性的自噬抑制剂与激活剂

　　●　快速量化原生异质性细胞群中的自噬

　　●　选择性地全方位染色，可检测和区分自噬流（autophagic flux）和自噬溶酶体的积累

　　●　不染溶酶体，减少其他染料的背景干扰

　　●　便于高通量筛选自噬激活剂和抑制剂

◆原理

　　该产品所用探针是阳离子两亲性示踪剂（CAT）染料，以类似诱导磷脂药物的方式迅速进入到细胞中。染料的功能基团能够选择性标记与自噬通路相关的小泡，而不会在溶酶体中聚集。

◆应用

CYTO-ID^® 自噬检测试剂盒 2.0：http://www.boppard.cn/product/show/735.html

◆相关产品

Cyto-ID®检测试剂盒 Q&A

Q：Cyto-ID®自噬检测试剂与活细胞孵育的最长时间是多少(几分钟/几小时/几天)？
A：我们建议37℃孵育不超过30 分钟。很多实验表明，孵育1 小时不会伤害细胞。孵育超过1 小时，一些细胞看
上去状态不好。

Q：是否可用荧光显微镜观察3D 培养细胞的自噬？
A：我们没有用该染料染过3D 细胞，但是一小撮细胞是可以染色的。活体组织的话，由于染料的渗透性不好，不
能进入样本的内部，所以效果不理想。染白细胞是可以。

Q：用双重染料染在37℃孵育30 分钟后，细胞状态不好开始皱缩。有些细胞也从板上脱离，阳性对照和DMSO
对照没有观察到特异性染色。
A：对于比较敏感的细胞，优化流程如下：
1. 使用PBS/5%FBS 或者完全培养基作为assay buffer，最好不含酚红。
2. 如果细胞状态仍然不好，建议染色时间缩短至15~20 分钟。
3. 减少染料的浓度至1000×稀释。需要显微镜观察时延长荧光照射的时间。
4. 可能光褪色会导致信号模糊。建议使用封片剂防止光褪色。

Q：用流式检测的话，样品在用Cyto-ID®自噬检测试剂孵育后多久可用流式观察？操作手册是说30 分钟。是否可
以在检测之前短暂的将样品放置在冰上？
A：可以将样品放在冰上延长孵育的时间。一般的经验法则，尽可能缩短活细胞与染料的结合。缺氧，低温或者其
他应激状态也可以诱导自噬。

Q：GFP-CertifiedTM 细胞凋亡/坏死检测试剂盒（ENZ-51002），Cyto-ID®自噬检测试剂盒(ENZ-51031)是否可以
同时染色，检测细胞凋亡，坏死和自噬？
A：GFP-CertifiedTM 细胞凋亡/坏死检测试剂盒（ENZ-51002）和Cyto-ID®检测试剂盒(ENZ-51031)不能配合使
用，因为细胞凋亡和自噬检测的染料都发绿色荧光。这两个试剂盒可以平行使用。比如，用十字孢碱
（Staurosporine）处理细胞，一部分样品用GFP-CertifiedTM 细胞凋亡/坏死检测试剂盒检测，一部分样品用
Cyto-ID®检测试剂盒检测。

Q：Cyto-ID®自噬检测试剂盒(ENZ-51031) 的Cyto-ID®自噬检测试剂能否与ProteoStat®蛋白聚集检测
（ENZ-51023）的ProteoStat®检测试剂联合使用？
A：Cyto-ID®绿色自噬检测试剂与ProteoStat®检测试剂发的荧光颜色不同，他们可用于同一实验，客户需要优化
实验流程。

Q：Cyto-ID®自噬检测试剂的原理是什么？
A：染料是申请专利的。该染料是阳离子两性示踪剂（CAT）染料，跟很多阳离子化合物一样可以快速进入细胞。
染料穿过细胞膜双分子层后被动扩散，不需要蛋白结合或者转运子活性。可滴定部分的筛选使的染料不会再溶
酶体聚集，能够标记自噬通道相关的液泡。另外，染料的发射光密度增强后可以标记自噬液泡相关片层膜结构。
我们可以提供染料染自噬液泡的相关数据。
诱导自噬溶酶体，比如用氯喹或者baflimyocin 处理后，会导致荧光变亮（250~300%）。雷帕霉素诱导自
噬后能够用3-MA 抑制。在EBSS 中氨基酸缺乏会导致1 小时后在自噬液泡中染料的积累，这种积累是可逆的
（补充营养后即可恢复）。我们有一系列用于研究自噬的化合物（Screen-Well®Autophagy library）

Q：与单丹磺酰尸胺(MDC)相比，Cyto-ID®自噬检测试剂有哪些优势？
A：MDC 需要365nm UV 荧光照明，与通常流式配置的488n 激发源不兼容。Cyto-ID®自噬染料是488nm激发
波长，绿色的发射荧光，可以高亮自噬通道不同的液泡成分。需要注意的是，与lysomotrophic 染料不同，
MDC，LysoTrackerTMRed，吖啶橙主要是染溶酶体的，Cyto-ID®自噬染料只能染一点点溶酶体，主要是染自
噬溶酶体和早期自噬部分的选择标记物。

Q：与GFP-LC3 转染细胞相比，Cyto-ID®自噬检测试剂盒(ENZ-51031)的优势是什么？
A：Cyto-ID®检测试剂盒(ENZ-51031)的最大优势是不用转染细胞。转染很短暂，不同实验需要再购买相关试剂。
由于细胞过表达，会导致GFP-LC3 聚集，结果呈假阳性。转染效率低于100%，有些细胞不会表达必要的GFP
组件。该试剂盒检测的自噬，细胞群的一致性会更好。不需要每个细胞系都进行转染。也可以标记原代细胞。
原代细胞不容易进行转染。

Q：有哪些细胞系或者原代细胞可用Cyto-ID®自噬检测试剂？
A：用该试剂盒见检测过的代表性的细胞和细胞系包括肝细胞，SK-N-SH 神经瘤细胞，CHO 细胞，骨肉瘤来源细
胞，黑素留细胞，乳腺癌细胞，宫颈癌细胞，卵巢癌细胞，B 淋巴瘤细胞，结肠癌细胞，HepG 细胞，Jurkat T
细胞，牛大动脉内皮细胞（BAEC）。

Q：Cyto-ID®自噬检测试剂能否与其他染料配合使用，区分细胞核内体和自噬内涵体？
A：Cyto-ID®红色长期示踪剂染料（ENZ-51037）能够染细胞质膜，通过细胞内吞存在于细胞中。Cyto-ID®红色
染料与Cyto-ID®自噬检测试剂能够高亮异体吞噬vs 自噬。也可以Cyto-ID®自噬检测试剂与Lyso-ID®红色染
料或者LysoTrackerTM 红色染料进行活细胞染色，荧光显微镜观察。

Q：Cyto-ID®自噬检测试剂的稳定性如何？
A：荧光染料会发生光褪色，但是这种染料比荧光素更稳定。按照操作手册出来荧光染料。

Q：Cyto-ID®自噬检测试剂盒染色后是否可固定细胞？
A：在操作手册中提供了固定细胞的流程。不推荐用表面活性剂给细胞打孔，定位的荧光染料丢失。

Q：活细胞检测时，荧光信号强度可维持多长时间？
A：细胞诱导后发生自噬，在分析前染色。诱导时间少于1 小时或者诱导几天，诱导时间取决于自噬诱导剂。我们
没有在活细胞上进行长时间的染色，有客户发现可以至少染色24 小时。

参考文献

A comparison of strategies for immortalizing mouse embryonic fibroblasts: M.M. St. Amad, et al.; J. Biol. Methods 3, e41 (2016), Application(s): Detected autophagy induction in SV40 transformed versus serially passed MEFs, 全文

Astemizole-Histamine induces Beclin-1-independent autophagy by targeting p53-dependent crosstalk between autophagy and apoptosis: R. Jakhar, et al.; Cancer Lett. 372, 89 (2016), Application(s): Flow cytometry analysis, 摘要;

Atg5 Is Essential for the Development and Survival of Innate Lymphocytes: T.E. O'Sullivan, et al.; Cell Rep. 15, 1910 (2016), Application(s): Liver lymphocytes were harvested and stained with cell surface antibodies and then incubated with 1:400 Cyto-ID Autophagy Detection Reagent, 摘要; 全文

Autophagy-related gene 5 and Wnt5 signaling pathway requires differentiation of embryonic stem cells into odontoblast-like cells: N. Ozeki, et al.; Exp. Cell Res. 341, 92 (2016), Application(s): Autophagy flux, 摘要;

DHA-induced stress response in human colon cancer cells-focus on oxidative stress and autophagy: K. Pettersen, et al.; Free Radic. Biol. Med. 90, 158 (2016), Application(s):Autophagy determined by flow cytometry, 摘要;

Diosgenin induces ROS-dependent autophagy and cytotoxicity via mTOR signaling pathway in chronic myeloid leukemia cells: S. Jiang, et al.; Phytomedicine 23, 243 (2016),Application(s): Confocal immunofluorescence, 摘要;

Efavirenz causes oxidative stress, endoplasmic reticulum stress, and autophagy in endothelial cells: M. Weiss, et al.; Cardiovasc. Toxicol. 16, 90 (2016), 摘要;

MiR-193b promotes autophagy and non-apoptotic cell death in oesophageal cancer cells: M.J. Nyhan, et al.; BMC Cancer 16, 101 (2016), Application(s): Assay used to stain live cells, 摘要; 全文

(Pro)renin receptor regulates autophagy and apoptosis in podocytes exposed to high glucose: C. Li, et al.; Am. J. Physiol. Endocrinol. Metab. 309, E302 (2015), Application(s):Confocal microscopy using mouse podocytes, 摘要;

A rapid and high content assay that measures cyto-ID-stained autophagic compartments and estimates autophagy flux with potential clinical applications: S. Guo, et al.; Autophagy11, 560 (2015), Application(s): Fluorescent detection, Microplate Reader , 摘要; 全文

A Systems Approach Identifies Essential FOXO3 Functions at Key Steps of Terminal Erythropoiesis: R. Liang, et al.; PLoS Genet. 11, e1005526 (2015), Application(s):Autophagy flux measured by flow cytometry , 摘要; 全文

ABT-888 enhances cytotoxic effects of temozolomide independent of MGMT status in serum free cultured glioma cells: R.K. Balvers, et al.; J. Transl. Med. 13, 74 (2015),Application(s): Assay, 摘要; 全文

Activation of autophagy in response to nanosecond pulsed electric field exposure: J.C. Ullery, et al.; Biochem. Biophys. Res. Commun. 458, 411 (2015), Application(s):Fluorescence microscopy using U937 monocyte and CHO-K1 cell lines, 摘要;

Aflatoxin biosynthesis is a novel source of reactive oxygen species-a potential redox signal to initiate resistance to oxidative stress?: L.V. Roze, et al.; Toxins (Basel). 7, 1411 (2015),Application(s): Assay, 摘要; 全文

Alisertib induces cell cycle arrest and autophagy and suppresses epithelial-to-mesenchymal transition involving PI3K/Akt/mTOR and sirtuin 1-mediated signaling pathways in human pancreatic cancer cells: F. Wang, et al.; Drug Des. Devel. Ther. 9, 575 (2015), Application(s): Flow cytometry using PANC-1 and BxPC-3 pancreatic cancer cell lines, 摘要; 全文

Alisertib, an Aurora kinase A inhibitor, induces apoptosis and autophagy but inhibits epithelial to mesenchymal transition in human epithelial ovarian cancer: Y.H. Ding, et al.; Drug Des. Devel. Ther. 9, 425 (2015), Application(s): Flow cytometry using SKOV3 and OVCAR-4 ovarian cancer cell lines, 摘要; 全文

Andrographolide Analogue Induces Apoptosis and Autophagy Mediated Cell Death in U937 Cells by Inhibition of PI3K/Akt/mTOR Pathway: D. Kumar, et al.; PLoS One 10, e0139657 (2015), Application(s): Flow cytometric analysis of Cyto-ID Green Detection Reagent , 摘要; 全文

Apoptotic Cell Death Induced by Resveratrol Is Partially Mediated by the Autophagy Pathway in Human Ovarian Cancer Cells: F. Lang, et al.; PLoS One 10, e0129196 (2015),Application(s): Live Cell Imaging, 摘要; 全文

Araguspongine C induces autophagic death in breast cancer cells through suppression of c-Met and HER2 receptor tyrosine kinase signaling: M.R. Akl, et al.; Mar. Drugs 13, 288 (2015), Application(s): Flow cytometry using BT-474 breast cancer cell line, 摘要; 全文

Autocrine VEGF maintains endothelial survival through regulation of metabolism and autophagy: C.K. Domigan, et al.; J. Cell. Sci. 128, 2236 (2015), 摘要;

Autophagy is activated in systemic lupus erythematosus and required for plasmablast development: A.J. Clarke, et al.; Ann. Rheum. Dis. 74, 912 (2015), 摘要; 全文

Autophagy limits proliferation and glycolytic metabolism in acute myeloid leukemia: A.S. Watson, et al.; Cell Death Discov. 1, 15008 (2015), Application(s): CytoID assay in human and mouse HSCs, 摘要; 全文

Baicalin inhibits autophagy induced by influenza A virus H3N2: H.Y. Zhu, et al.; Antiviral Res. 113, 62 (2015), Application(s): Fluorescence microscopy using A549 human lung cancer cell line, 摘要;

Bardoxolone methyl induces apoptosis and autophagy and inhibits epithelial-to-mesenchymal transition and stemness in esophageal squamous cancer cells: Y.Y. Wang, et al.; Drug Des. Devel. Ther. 9, 993 (2015), Application(s): Flow Cytometry, 摘要; 全文

Cell-penetrating peptide derived from human eosinophil cationic protein inhibits mite allergen Der p 2 induced inflammasome activation: S.J. Yu, et al.; PLoS One 10, e0121393 (2015), Application(s): Flow cytometry of THP-1 leukemia cell line, 摘要; 全文

Chemoproteomics Reveals Novel Protein and Lipid Kinase Targets of Clinical CDK4/6 Inhibitors in Lung Cancer: N.J. Sumi, et al.; ACS Chem. Biol. 10, 2680 (2015),Application(s): Quantification of autophagosomes, 摘要;

Circulating hemocytes from larvae of the Japanese rhinoceros beetle Allomyrina dichotoma (Linnaeus) (Coleoptera: Scarabaeidae) and the cellular immune response to microorganisms: S. Hwang, et al.; PLoS One 10, e0128519 (2015), Application(s):Fluorescence microscopy using hemocytes from Japanese rhinoceros beetle Allomyrina dichotoma larvae, 摘要; 全文

Citreoviridin induces ROS-dependent autophagic cell death in human liver HepG2 cells: Y.N. Liu, et al.; Toxicon. 95, 30 (2015), Application(s): Fluorescence microscopy using HepG2 cell line, 摘要;

Clozapine induces autophagic cell death in non-small cell lung cancer cells: Y.C. Yin, et al.; Cell. Physiol. Biochem. 35, 945 (2015), 摘要;

Coffee and caffeine potentiate the antiamyloidogenic activity of melatonin via inhibition of Aβ oligomerization and modulation of the Tau-mediated pathway in N2a/APP cells: L.F. Zhang, et al.; Drug Des. Devel. Ther. 9, 241 (2015), Application(s): Flow Cytometry,摘要; 全文

Combination of the mTOR inhibitor RAD001 with temozolomide and radiation effectively inhibits the growth of glioblastoma cells in culture: H. Burckel, et al.; Oncol. Rep. 33, 471 (2015), 摘要;

Danusertib Induces Apoptosis, Cell Cycle Arrest, and Autophagy but Inhibits Epithelial to Mesenchymal Transition Involving PI3K/Akt/mTOR Signaling Pathway in Human Ovarian Cancer Cells: D. Zi, et al.; Int. J. Mol. Sci. 16, 27228 (2015), Application(s): Confocal fluorescence microscopy, 摘要; 全文

Danusertib, a potent pan-Aurora kinase and ABL kinase inhibitor, induces cell cycle arrest and programmed cell death and inhibits epithelial to mesenchymal transition involving the PI3K/Akt/mTOR-mediated signaling pathway in human gastric cancer AGS and NCI-N78 cells: C.X. Yuan, et al.; Drug Des. Devel. Ther. 9, 1293 (2015), Application(s): Flow cytometry using AGS and NCI-N78 gastric cancer cell lines, 摘要; 全文

Defective autophagy in vascular smooth muscle cells alters contractility and Ca²⁺ homeostasis in mice: C.F. Michiels, et al.; Am. J. Physiol. Heart Circ. Physiol. 308, H557 (2015), 摘要;

Effects of cyclodextrins on GM1‐gangliosides in fibroblasts from GM1‐gangliosidosis patients: Y. Maeda, et al.; J. Pharm. Pharmacol. 67, 1133 (2015), 摘要;

Endurance exercise training induces fat depot-specific differences in basal autophagic activity: G. Tanaka, et al.; Biochem. Biophys. Res. Commun. 466, 512 (2015),Application(s): Detect the formation of autophagosomes, 摘要;

Erbin is a novel substrate of the Sag-βTrCP E3 ligase that regulates KrasG12D-induced skin tumorigenesis: C.M. Xie, et al.; J. Cell. Biol. 209, 721 (2015), 摘要;

Evaluation of Antitumor Effects of Folate-Conjugated Methyl-β-cyclodextrin in Melanoma: K. Motoyama, et al.; Biol. Pharm. Bull. 38, 374 (2015), Application(s): Fluorescence Microscopy, 摘要; 全文

Exchange protein directly activated by cAMP 1 promotes autophagy during cardiomyocyte hypertrophy: A.C. Laurent, et al.; Cardiovasc. Res. 105, 55 (2015), Application(s):Fluorescence microscopy using rat neonatal ventricular myocytes, 摘要;

Glutathione-S-transferase omega 1 (GSTO1-1) acts as mediator of signaling pathways involved in aflatoxin B1-induced apoptosis-autophagy crosstalk in macrophages: S. Paul, et al.; Free Radic. Biol. Med. 89, 1218 (2015), Application(s): Determination of autophagy with immunocytochemistry , 摘要;

GMI, an immunomodulatory protein from Ganoderma microsporum, potentiates cisplatin-induced apoptosis via autophagy in lung cancer cells: I.L. Hsin, et al.; Mol. Pharm. 12, 1534 (2015), 摘要;

Induction of apoptosis and autophagy via sirtuin1- and PI3K/Akt/mTOR-mediated pathways by plumbagin in human prostate cancer cells: Z.W. Zhou, et al.; Drug Des. Devel. Ther. 9, 1511 (2015), Application(s): Assay, 摘要; 全文

Induction of autophagy is a key component of all-trans-retinoic acid-induced differentiation in leukemia cells and a potential target for pharmacologic modulation: N. Orfali, et al.; Exp. Hematol. 43, 781 (2015), Application(s): Flow cytometry analysis of NB4 and HL60 promyelocytic leukemia cell lines, 摘要;

Inhibition of Autophagy Potentiated the Antitumor Effect of Nedaplatin in Cisplatin-Resistant Nasopharyngeal Carcinoma Cells: Z. Liu, et al. ; PLoS One 10, e0135236 (2015),Application(s): Cell culture, 摘要; 全文

Inhibition of mitotic Aurora kinase A by alisertib induces apoptosis and autophagy of human gastric cancer AGS and NCI-N78 cells: C.X. Yuan, et al.; Drug Des. Devel. Ther. 9, 487 (2015), Application(s): Flow cytometry using AGS and NCI-N78 gastric cancer cell lines, 摘要; 全文

Interferon Regulatory Factor-1 signaling regulates the switch between autophagy and apoptosis to determine breast cancer cell fate: J.L. Schwartz-Roberts, et al.; Cancer Res.75, 1046 (2015), 摘要;

Interplay of Oxidative Stress and Autophagy in PAMAM Dendrimers-Induced Neuronal Cell Death : Y. Li, et al.; Theranostics 5, 1363 (2015), Application(s): Confocal fluorescence assay, 摘要; 全文

Invariant NKT cells require autophagy to coordinate proliferation and survival signals during differentiation: B. Pei, et al.; J. Immunol. 194, 5872 (2015), 摘要;

Involvement of fish signal transducer and activator of transcription 3 (STAT3) in nodavirus infection induced cell death: Y. Huang, et al.; Fish Shellfish Immunol. 43, 241 (2015),Application(s): Fluorescence microscopy of Grouper (fish) brain cells, 摘要;

Is the autophagy a friend or foe in the silver nanoparticles associated radiotherapy for glioma?: H. Wu, et al.; Biomaterials 62, 47 (2015), Application(s): Fluorescence microscopy using U251 human glioma cell line, 摘要;

Kaposi's sarcoma-associated herpesvirus induces Nrf2 activation in latently infected endothelial cells through SQSTM1 phosphorylation and interaction with polyubiquitinated Keap1: O. Gjyshi, et al.; J. Virol. 89, 2268 (2015), 摘要;

KLF4-SQSTM1/p62-associated prosurvival autophagy contributes to carfilzomib resistance in multiple myeloma models: I. Riz, et al.; Oncotarget 6, 17814 (2015), Application(s):FACS, 摘要; 全文

Lithium modulates autophagy in esophageal and colorectal cancer cells and enhances the efficacy of therapeutic agents in vitro and in vivo: T.R. O'Donovan, et al.; PLoS One 10, e0134676 (2015), Application(s): Flow cytometry analysis using human esophageal and murine colon cancer cell lines, 摘要; 全文

Methicillin-Resistant Staphylococcus aureus Adaptation to Human Keratinocytes: G. Soong, et al.; MBio. 6, e00289-15 (2015), Application(s): Assay, 摘要; 全文

Mevalonate pathway regulates cell size homeostasis and proteostasis through autophagy: T.P. Miettinen, et al.; Cell Rep. 13, 2610 (2015), Application(s): Flow cytometry analysis of autophagy using Jurkat, U2OS, Kc167 and HUVEC cells, 摘要;

MiR-29b replacement inhibits proteasomes and disrupts aggresome+autophagosome formation to enhance the antimyeloma benefit of bortezomib: S. Jagannathan, et al.; Leukemia 29, 727 (2015), Application(s): Detection of autophagy by fluorescence microscopy in multiple myeloma cell lines, 摘要; 全文

Molecular chaperone GRP78 enhances aggresome delivery to autophagosomes to promote drug resistance in multiple myeloma: M.A. Abdel Malek, et al.; Oncotarget 6, 3098 (2015), Application(s): Confocal Microscopy, 摘要; 全文

Molecular cloning and characterization of autophagy-related gene TmATG8 in Listeria-invaded hemocytes of Tenebrio molitor: H. Tindwa, et al.; Dev. Comp. Immunol. 51, 88 (2015), Application(s): Fluorescence microscopy using hemocytes from Tenebrio molitor larvae, 摘要;

Molecular pathway of near-infrared laser phototoxicity involves ATF-4 orchestrated ER stress: I. Khan, et al.; Sci. Rep. 5, 10581 (2015), Application(s): Fluorescence microscopy autophagy assay, 摘要; 全文

N-Myc and STAT Interactor regulates autophagy and chemosensitivity in breast cancer cells: B.J. Metge, et al.; Sci. Rep. 5, 11995 (2015), Application(s): Fluorescent detection,摘要; 全文

Novel autophagy inducers lentztrehaloses A, B and C: S.I. Wada, et al.; J. Antibiot. (Tokyo)68, 521 (2015), Application(s): Fluorescence microscopy using Mewo melanoma and OVK18 ovarian cancer cell lines, 摘要;

Novel small-molecule SIRT1 inhibitors induce cell death in adult T-cell leukaemia cells: T. Kozako, et al.; Sci. Rep. 5, 11345 (2015), Application(s): Flow cytometry using a variety of cancer cell lines, 摘要; 全文

Novel targeting of PEGylated liposomes for codelivery of TGF-β1 siRNA and four antitubercular drugs to human macrophages for the treatment of mycobacterial infection: a quantitative proteomic study: N. Niu, et al. ; Drug Des. Devel. Ther. 9, 4441 (2015),Application(s): Autophagy of human macrophages by flow cytometry, 摘要;

Paraptosis cell death induction by the thiamine analog benfotiamine in leukemia cells: N. Sugimori, et al.; PLoS One 10, e0120709 (2015), Application(s): Flow cytometry using HL60 leukemia cell line, 摘要; 全文

Plumbagin induces G2/M arrest, apoptosis, and autophagy via p38 MAPK- and PI3K/Akt/mTOR-mediated pathways in human tongue squamous cell carcinoma cells: S.T. Pan, et al.; Drug Des. Devel. Ther. 9, 1601 (2015), Application(s): Assay, 摘要; 全文

Pro-apoptotic and pro-autophagic effects of the Aurora kinase A inhibitor alisertib (MLN8237) on human osteosarcoma U-2 OS and MG-63 cells through the activation of mitochondria-mediated pathway and inhibition of p38 MAPK/PI3K/Akt/mTOR signaling pathway: N.K. Niu, et al.; Drug Des. Devel. Ther. 9, 1555 (2015), Application(s): Assay,摘要; 全文

Reduced FoxO3a expression causes low autophagy in idiopathic pulmonary fibrosis fibroblasts on collagen matrix: J. Im, et al.; Am. J. Physiol. Lung Cell. Mol. Physiol. 309, L552 (2015), 摘要;

S-Adenosyl-L-methionine-competitive inhibitors of the histone methyltransferase EZH2 induce autophagy and enhance drug sensitivity in cancer cells: T.P. Liu, et al.; Anticancer Drugs 26, 139 (2015), Application(s): Fluorescence microscopy using MDA-MB-231 breast cancer cell line, 摘要; 全文

Schisandrin B inhibits cell growth and induces cellular apoptosis and autophagy in mouse hepatocytes and macrophages: implications for its hepatotoxicity: Y. Zhang, et al.; Drug Des. Devel. Ther. 9, 2001 (2015), Application(s): Flow cytometry using AML-12 hepatocyte and RAW 264.7 leukemia cell lines, 摘要; 全文

Src/STAT3-dependent heme oxygenase-1 induction mediates chemoresistance of breast cancer cells to doxorubicin by promoting autophagy: Q. Tan, et al.; Cancer Sci. 106, 1023 (2015), 摘要;

The CCL2 chemokine is a negative regulator of autophagy and necrosis in luminal B breast cancer cells: W.B. Fang, et al.; Breast Cancer Res. Treat. 150, 309 (2015), 摘要;

The investigational Aurora kinase A inhibitor alisertib (MLN8237) induces cell cycle G2/M arrest, apoptosis, and autophagy via p38 MAPK and Akt/mTOR signaling pathways in human breast cancer cells: J.P. Li, et al.; Drug Des. Devel. Ther. 9, 1627 (2015),Application(s): Assay, 摘要; 全文

The pan-inhibitor of Aurora kinases danusertib induces apoptosis and autophagy and suppresses epithelial-to-mesenchymal transition in human breast cancer cells: J.P. Li, et al.; Drug Des. Devel. Ther. 9, 1027 (2015), Application(s): Assay, 摘要; 全文

The role of autophagy in the cytotoxicity induced by recombinant human arginase in laryngeal squamous cell carcinoma: C. Lin, et al.; Appl. Microbiol. Biotechnol. 99, 8487 (2015), 摘要;

A Novel CXCR3-B Chemokine Receptor-induced Growth-inhibitory Signal in Cancer Cells Is Mediated through the Regulation of Bach-1 Protein and Nrf2 Protein Nuclear Translocation : M. Balan & S. Pal; J. Biol. Chem. 289, 3126 (2014), Application(s): Monitor autophagy in MCF-7 and T47D breast cancer cells by flow cytometry and fluorescence microscopy, 摘要;

Adaptive responses to glucose restriction enhance cell survival, antioxidant capability, and autophagy of the protozoan parasite Trichomonas vaginalis: K.Y. Huang, et al.; Biochim. Biophys. Acta. 1840, 53 (2014), 摘要;

Autophagy in the brain of neonates following hypoxia-ischemia shows sex-and region-specific effects: S.N. Weis, et al.; Neuroscience 256, 201 (2014), 摘要;

Cannabinoid-induced autophagy regulates suppressor of cytokine signaling-3 in intestinal epithelium: L.C. Koay, et al.; Am. J. Physiol. Gastrointest. Liver Physiol. 307, G140 (2014),Application(s): Detection of autophagy in human colonic epithelial cell line Caco-2 by Confocal imaging, 摘要; 全文

Caveolin-1 Is a Critical Determinant of Autophagy, Metabolic Switching, and Oxidative Stress in Vascular Endothelium: T. Shiroto, et al.; PLoS One 9, e87871 (2014), 摘要;全文

Connective tissue diseases: How do autoreactive B cells survive in SLE-autophagy?: N.J. Bernard; Nat. Rev. Rheumatol. 10, 128 (2014), (Review), 摘要;

Defective Autophagosome Trafficking Contributes to Impaired Autophagic Flux in Coronary Arterial Myocytes Lacking CD38 Gene: Y. Zhang, et al.; Cardiovasc. Res. 102, 68 (2014),摘要;

Defects in mitochondrial clearance predispose human monocytes to interleukin-1β hyper-secretion: R. van der Burgh, et al.; J. Biol. Chem. 289, 5000 (2014), 摘要; 全文

Early biomarkers of response to carfilzomib in multiple myeloma (MM): Modulation of CXCR4 and induction of autophagy: M. Bhutani, et al.; J. Clin. Oncol. 32, e19572 (2014),Application(s): Quantification of autophagy in malignant plasma cells from bone marrow aspirates by flow cytometry with the Cyto-ID autophagy detection kit,

Enhancement of dynein-mediated autophagosome trafficking and autophagy maturation by ROS in mouse coronary arterial myocytes: M. Xu, et al.; J. Cell. Mol. Med. 18, 2165 (2014), 摘要; 全文

Flow Cytometric Analysis of Autophagic Activity with Cyto-ID Staining in Primary Cells: M. Stankov, et al.; Bio-Protocol (2014), Application(s): FC in primary BMDCs, 全文

High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell-Derived Cells: O. Sirenko, et al.; Assay Drug Dev. Technol. 12, 43 (2014), 摘要; 全文

Histone deacetylase inhibitors induce apoptosis in myeloid leukemia by suppressing autophagy: M.V. Stankov, et al.; Leukemia 28, 577 (2014), 摘要;

Histone deacetylase inhibitors potentiate VSV oncolysis in prostate cancer cells by modulating NF-κB dependent autophagy: L. Shulak, et al.; J. Virol. 88, 2927 (2014),摘要;

In vitro and in vivo characterization of porcine acellular dermal matrix for gingival augmentation procedures: A.M. Pabst, et al.; J. Periodontal. Res. 49, 371 (2014), 摘要;

Inhibition of Autophagic Flux by Salinomycin Results in Anti-Cancer Effect in Hepatocellular Carcinoma Cells: J. Klose, et al.; PLoS One 9, e95970 (2014),Application(s): Autophagy detection in human hepatocellular carcinoma , 摘要; 全文

Inhibition of stress induced premature senescence in presenilin-1 mutated cells with water soluble Coenzyme Q10: D. Ma, et al.; Mitochondrion 17C, 106 (2014), Application(s):Autophagic vacuoles in Alzheimer's Disease fibroblasts detected with CytoID® Green Autophagy Detection kit, 摘要;

Involvement of autophagy in recombinant human arginase-induced cell apoptosis and growth inhibition of malignant melanoma cells: Z. Wang, et al.; Appl. Microbiol. Biotechnol.98, 2485 (2014), 摘要;

MiR-216a: a link between endothelial dysfunction and autophagy: R. Menghini, et al.; Cell Death Dis. 5, e1029 (2014), 摘要;

Novel estradiol analogue induces apoptosis and autophagy in esophageal carcinoma cells: E. Wolmarans, et al.; Cell. Mol. Biol. Lett. 19 , 98 (2014), Application(s): Autophagy detection in esophageal carcinoma SNO cell , 摘要;

Novel sorafenib-based structural analogues: in-vitro anticancer evaluation of t-MTUCB and t-AUCMB: A.T. Wecksler, et al.; Anticancer Drugs 25, 433 (2014), 摘要;

Photodynamic therapy with the novel photosensitizer chlorophyllin f induces apoptosis and autophagy in human bladder cancer cells: D. Lihuan, et al.; Lasers Surg. Med. 46, 319 (2014), 摘要;

Plumbagin induces apoptotic and autophagic cell death through inhibition of the PI3K/Akt/mTOR pathway in human non-small cell lung cancer cells: Y.C.Li, et al.; Cancer Lett. 344, 239 (2014), 摘要;

Potential of adenovirus-mediated REIC/Dkk-3 gene therapy for use in the treatment of pancreatic cancer: D. Uchida, et al.; J. Gastroenterol. Hepatol. 29, 973 (2014), 摘要;

Sirt1 modulates endoplasmic reticulum stress-induced autophagy in heart: A. Guilbert, et al.; Cardiovasc. Res. 103 (suppl 1), S13 (2014), Application(s): Evaluation of Autophagy in H9c2 cells, rat cardiomyoblasts by flow cytometry, 全文

STAT3 down regulates LC3 to inhibit autophagy and pancreatic cancer cell growth: J. Gong, et al.; Oncotarget 5, 2529 (2014), Application(s): Autophagic vacuole formation was detected by microscopy and autophagosome formation was determined by flow cytometry in human pancreatic cancer cells Capan-2, 摘要; 全文

T-Cell Autophagy Deficiency Increases Mortality and Suppresses Immune Responses after Sepsis: C.W. Lin, et al.; PLoS One 9, e102066 (2014), Application(s): Quantification of autophagosomes and autolysosomes staining in CD4+ and CD8+ cell population by flow cytometry , 摘要; 全文

Tetracyclines cause cell stress-dependent ATF4 activation and mTOR inhibition: A. Brüning, et al.; Exp. Cell Res. 320, 281 (2014), 摘要;

The core autophagy protein ATG4B is a potential biomarker and therapeutic target in CML stem/progenitor cells: K. Rothe, et al.; Blood 123, 3622 (2014), Application(s): Monitor autophagy flux in hematopoietic stem/progenitor cells, 摘要;

Androgen deprivation and androgen receptor competition by bicalutamide induce autophagy of hormone-resistant prostate cancer cells and confer resistance to apoptosis: B. Boutin, et al.; Prostate 73, 1090 (2013), Application(s): Measurement of autophagic flux in prostate cancer cells, 摘要;

Arenobufagin, a natural bufadienolide from toad vonem, induces apoptosis and autophagy in human hepatocellular carcinoma cells through inhibition of PI3K/Akt/mTOR pathway: D.M. Zhang, et al.; Carcinogenesis 34, 1331 (2013), Application(s): Autophagy detection in hepatocellular carcinoma, 摘要;

Autophagy Plays a Critical Role in ChLym-1-Induced Cytotoxicity of Non-Hodgkin's Lymphoma Cells: J. Fan, et al.; PLoS One. 8, e72478 (2013), 摘要; 全文

BCL-2 inhibitors sensitize therapy-resistant chronic lymphocytic leukemia cells to VSV oncolysis: S. Samuel, et al.; Mol. Ther. 21, 1413 (2013), 摘要;

Bleomycin exerts ambivalent antitumor immune effect by triggering both immunogenic cell death and proliferation of regulatory T cells: H. Bugaut, et al.; PLoS One 8, e65181 (2013),Application(s): Measurement of autophagy by flow cytometry and fluorescence microscopy, 摘要; 全文

Celecoxib enhances radiosensitivity of hypoxic glioblastoma cells through endoplasmic reticulum stress: K. Suzuki, et al.; Neuro. Oncol. 15, 1186 (2013), 摘要;

Chloroquine Engages the Immune System to Eradicate Irradiated Breast Tumors in Mice: J.A. Ratikan, et al.; Int. J. Radiat. Oncol. Biol. Phys. 87, 761 (2013), 摘要;

Dietary Resveratrol Prevents Development of High-Grade Prostatic Intraepithelial Neoplastic Lesions: Involvement of SIRT1/S6K Axis: G. Li, et al.; Cancer Prev. Res 6, 27 (2013), Application(s): Effects of Resveratrol on prostate tumorigenesis, 摘要;

Enhancement of autophagy by simvastatin through inhibition of Rac1-mTOR signaling pathway in coronary arterial myocytes: Y.M. Wei, et al.; Cell. Physiol. Biochem. 31, 925 (2013), 摘要; 全文

GX15-070 (obatoclax) induces apoptosis and inhibits cathepsin D and L mediated autophagosomal lysis in antiestrogen resistant breast cancer cells: J.L. Schwartz-Roberts, et al.; Mol. Cancer Ther. 12, 448 (2013), Application(s): Autophagy detection in breast cancer cells, 摘要;

Hydroxychloroquine preferentially induces apoptosis of CD45RO+ effector T cells by inhibiting autophagy: A possible mechanism for therapeutic modulation of T cells: J. van Loodregt, et al.; J. Allergy Clin. Immunol. 131, 1443 (2013), Application(s): Detection of autophagy in CD4+ T cells and PBMC by flow cytometry , 摘要; 全文

Interactions between autophagic and endo-lysosomal markers in endothelial cells: C.L. Oeste, et al.; Histochem. Cell. Biol. 139, 659 (2013), 摘要;

Involvement of cholesterol depletion from lipid rafts in apoptosis induced by methyl-β-cyclodextrin: R. Onodera, et al.; Int. J. Pharm. 452, 116 (2013), Application(s):Measurement of autophagy by fluorescence microscopy, 摘要;

ISG15 deregulates autophagy in genotoxin-treated ataxia telangiectasia cells: S.D. Desai, et al.; J. Biol. Chem. 288, 2388 (2013), Application(s): Fluorescence microscopy using Ataxia Telangiectasia cells, 摘要; 全文

Lysosomal basification and decreased autophagic flux in oxidatively stressed trabecular meshwork cells: Implications for glaucoma pathogenesis: K. Porter, et al.; Autophagy 9, 581 (2013), Application(s): Autophagy detection by flow cytometry in porcine TM cells,摘要; 全文

Nelfinavir and bortezomib inhibit mTOR activity via ATF4-mediated sestrin-2 regulation: A. Brüning; Mol. Oncol. 7, 1012 (2013), 摘要;

Recombinant human arginase induced caspase-dependent apoptosis and autophagy in non-Hodgkin's lymphoma cells: X. Zeng, et al.; Cell Death Dis. 4, e840 (2013), 摘要;全文

Regulation of autophagic flux by dynein-mediated autophagosomes trafficking in mouse coronary arterial myocytes: M. Xu, et al.; Biochim. Biophys. Acta. 1833, 3228 (2013),摘要;

Renal cancer-selective Englerin A induces multiple mechanisms of cell death and autophagy: R.T. Williams, et al.; J. Exp. Clin. Cancer Res. 32, 57 (2013), Application(s):Flow cytometry and immunofluorescence of a human kidney carcinoma cell line, 摘要;全文

Saxifragifolin D induces the interplay between apoptosis and autophagy in breast cancer cells through ROS-dependent endoplasmic reticulum stress: J.M. Shi, et al.; Biochem. Pharmacol. 85, 913 (2013), Application(s): Autophagy detection by flow cytometry in breast cancer cells, 摘要;

Suppression of autophagy enhanced growth inhibition and apoptosis of interferon-β in human glioma cells: Y. Li, et al.; Mol. Neurobiol. 47, 1000 (2013), 摘要;

Survival and death strategies in glioma cells: autophagy, senescence and apoptosis triggered by a single type of temozolomide-induced DNA damage: A.V. Knizhnik, et al.; PLoS One 8, e55665 (2013), Application(s): Autophagy detection by flow cytometry in glioma cells, 摘要; 全文

The effect of Zhangfei on the unfolded protein response and growth of cells derived from canine and human osteosarcomas: T. Bergeron, et al.; Vet. Comp. Oncol. 11, 140 (2013),Application(s): Detection of autophagy in human and canine osteosarcoma, 摘要;

The mTOR inhibitor RAD001 potentiates autophagic cell death induced by temozolomide in a glioblastoma cell line: E. Josset, et al.; Anticancer Res. 33, 1845 (2013), 摘要;

Therapeutic Combination of Nanoliposomal Safingol and Nanoliposomal Ceramide for Acute Myeloid Leukemia: T.J. Brown, et al.; J. Leuk. 1, 110 (2013), Application(s):Detection of autophagy by flow cytometry in Human HL-60 , HL-60/VCR, and murine C1498 cells, 全文

Type I interferons induce autophagy in certain human cancer cell lines: H. Schmeisser, et al.; Autophagy 9, 683 (2013), Application(s): Autophagy detection in type I interferon-treated human cancer cell lines, 摘要;

A novel image-based cytometry method for autophagy detection in living cells: L.L. Chan, et al.; Autophagy 8, 1371 (2012), 摘要; 全文

Apoptosis and autophagy have opposite roles on imatinib-induced K562 leukemia cell senescence: C. Drullion, et al.; Cell Death Dis. 3, e373 (2012), Application(s): Flow cytometry of human CML cells treated with Imatinib, 摘要; 全文

Counteracting autophagy overcomes resistance to everolimus in mantle cell lymphoma: L. Rosich, et al.; Clin. Cancer Res. 18, 5278 (2012), 摘要; 全文

Heme Oxygenase-1 Promotes Survival of Renal Cancer Cells through Modulation of Apoptosis-and Autophagy-regulating Molecules: P. Banerjee, et al.; J. Biol. Chem. 287, 4962 (2012), Application(s): Detection of autophagy in human renal cancer cells, 摘要;

Inhibition of monocarboxylate transporter 2 induces senescence-associated mitochondrial dysfunction and suppresses progression of colorectal malignancies in vivo: I. Lee, et al.; Mol. Cancer Ther. 11, 2342 (2012), 摘要; 全文

Mechanism for the induction of cell death in ONS-76 medulloblastoma cells by Zhangfei/CREB-ZF: T.W. Bodnarchuk, et al.; J. Neurooncol. 109, 485 (2012),Application(s): Detection of autophagy in medulloblastoma cells, 摘要;

Mitochondrial metabolism in Parkinson's disease impairs quality control autophagy by hampering microtubule-dependent traffic: D.M. Arduíno, et al.; Hum. Mol. Genet. 21, 4680 (2012), 摘要; 全文

Proteasome inhibition by quercetin triggers macroautophagy and blocks mTor activity: A.K. Klappan, et al.; Histochem. Cell Biol. 137, 25 (2012), 摘要;

Reovirus as a viable therapeutic option for the treatment of multiple myeloma: C.M. Thirukkumaran, et al.; Clin. Cancer Res. 18, 4962 (2012), Application(s): Detection of autophagy in human myeloma cell lines and ex vivo tumor specimens, 摘要;

Src inhibition with saracatinib reverses fulvestrant resistance in ER-positive ovarian cancer models in vitro and in vivo: F.A. Simpkins, et al.; Clin. Cancer Res. 18, 5911 (2012),Application(s): Detection of autophagy in human ovarian cancer cells and xenografts,摘要;

FoxM1 knockdown sensitizes human cancer cells to proteasome inhibitor-induced apoptosis but not to autophagy: B. Pandit, et al.; Cell Cycle 10, 3269 (2011), Application(s):Flow cytometry using human cancer cells, 摘要; 全文

Monitoring of autophagy in Chinese hamster ovary cells using flow cytometry: J.S. Lee, et al.; Methods 56(3), 375 (2011), 摘要;

Selective anticancer activity of a hexapeptide with sequence homology to a non-kinase domain of Cyclin Dependent Kinase 4: H.M. Warenius, et al.; Mol. Cancer 10, 72 (2011),摘要;

Silibinin triggers apoptotic signaling pathways and autophagic survival response in human colon adenocarcinoma cells and their derived metastatic cells: H. Kauntz, et al.; Apoptosis16, 1042 (2011), 摘要;

组织标本脱色试剂

deColorizing Solution 1

deColorizing Solution 2

本产品是由麻布大学的小泽秋沙先生和坂上元荣先生共同开发的对组织染色标本进行脱色的全新型试剂。对病理学领域中广泛使用的苏木精和伊红染色（HE染色）以及Masson三色染色（MT染色）等特殊染色进行脱色。

在对HE染色标本脱色时，只需使用deColorizing Solution 1。另外，MT染色标本脱色时需要使用deColorizing Solution 1和deColorizing Solution 2。

使用本产品进行脱色后，通过进行不同的染色，可以从一个组织标本中获得多个染色信息。

◆特点

●　可对HE染色和MT染色进行脱色

●　脱色后可再进行抗体染色或HE染色等。

●　操作简单，只需本产品浸泡即可。

◆应用实例

■使用小鼠脑（海马）标本进行HE染色时

使用deColorizing Solution 1进行脱色

■使用小鼠肝脏标本进行MT染色时

使用deColorizing Solution 1和2进行脱色

左至右：MT染色→脱色→HE染色

（数据提供：麻布大学 小泽秋沙先生，坂上元荣先生）

抗FGF1，单克隆抗体（mAb1）

Anti FGF1, Monoclonal Antibody (mAb1)

抗大鼠LC3，兔

Anti Rat LC3, Rabbit

分泌酶抑制剂

阿尔茨海默病研究相关试剂

　　β-淀粉样蛋白（Aβ）是阿尔茨海默病主要病理特征——老人斑的主要构成成分，是通过β分泌酶和γ分泌酶切断β-淀粉样蛋白前体蛋白质（APP）而产生的。因此，分泌酶抑制剂可作为β-淀粉样蛋白产生抑制剂进行研究

相关产品

β-分泌酶ELISA试剂盒

　　本产品是可高灵敏度检测β-淀粉样蛋白（1-40）和β-淀粉样蛋白（1-42）的Elisa试剂盒。不仅能检测组织提取液、培养上清和脑脊髓液，甚至连难以检测的血浆中样品，也可高灵敏度检测出来。

相关资料

阿尔兹海默症相关系列产品

S-100β ELISA试剂盒

可特异性检测出脑损伤标记S-100β产品

 S-100β 蛋白分子量为21KD，有两个亚基组成（α 亚基和β 亚基），在人体中这些亚基的组合及其定位是不同的。S-100ββ 主要存在于神经胶质细胞和雪旺细胞中，S-100αβ 主要存在于神经胶质细胞中，S-100αα主要存在于横纹肌、心脏和肾脏中。

S-100β 是一种钙结合蛋白，有研究证明脑脊液中的S-100β 是临床诊断中脑损伤的重要标志物。S-100β 与脑缺血、脑病理性出血、重度颅脑损伤造成的脑损伤密切相关，在阿尔茨海默、帕金森氏病的患者的血液中也能检测到S-100β 的含量升高。

该试剂盒用于定量检测小鼠、大鼠和人血浆和脑脊液样品中S-100β。

◆优点

●　高灵敏性、高再现性

●　样品无需太多

●　已准备需要的相关试剂

◆性能一览

◆试剂组成

1. Antibody Coated Plate————————– 1 Plate

2. S-100β Standard ——————————– 5 ng

3. Biotinylated Rabbit Anti Bovine S-100β ——- 11 ml

4. HRP Labeled Streptavidin Solution————- 11 ml

5. Substrate Buffer——————————— 26 ml

6. OPD Tablet————————————— 2 tablets

7. Stopping Solution——————————- 11 ml

8. Buffer Solution———————————- 25 ml

9. Washing Solution (concentrated)————— 50 ml

10. Adhesive Foil———————————– 4 sheets

◆测定原理

检量线

[1]　Fabrizio, M. et al.: Clinical Chemistry., 48, 2097 (2002).

[2]　Green, A.J. et al.: NeurosciLett., 235, 5 (1997).

[3]　Kinnari, S. et al.: Brain., 135, 3336 (2012).