Annexin V-FITC/PI细胞凋亡检测试剂盒是用FITC标记的Annexin V作为探针，来检测细胞早期凋亡的发生。

其检测原理为：在正常的活细胞中，磷脂酰丝氨酸（phosphotidylserine，PS）位于细胞膜的内侧，但在早期凋亡的细胞中，PS 从细胞膜的内侧翻转到细胞膜的表面，暴露在细胞外环境中。Annexin-Ⅴ（膜联蛋白-V）是一种分子量为35-36 kDa的Ca²⁺ 依赖性磷脂结合蛋白，能与PS高亲和力结合，可通过细胞外侧暴露的磷脂酰丝氨酸与凋亡早期细胞的胞膜结合。

另外，本试剂盒中还提供了碘化丙啶（Propidium Iodide，PI）用来区分存活的早期细胞和坏死或晚期凋亡细胞。PI是一种核酸染料，它不能透过正常细胞或早期凋亡细胞的完整的细胞膜，但可以透过凋亡晚期和坏死细胞的细胞膜而使细胞核染红。因此，将Annexin V与PI联合使用时，PI 则被排除在活细胞（Annexin V-/PI-）和早期凋亡细胞（Annexin V+/PI-）之外，而晚期凋亡细胞和坏死细胞同时被FITC 和PI 结合染色呈现双阳性（Annexin V+/PI+）。

本试剂盒可用于流式细胞仪、荧光显微镜进行检测。

检测便捷、准确

冰袋（wet ice）运输。-20℃避光保存，避免反复冻融，一年有效。

【注】：如果需要在短时间内多次重复使用，可以在4℃避光保存，半年有效。

Q：Annexin V 和 JC-1、Tunel 细胞凋亡检测的区别？

A: Annexin V 是检测细胞早期凋亡的试剂，JC-1 是检测细胞中期凋亡的试剂、Tunel 是检测细胞晚期凋亡的试剂。

Q：Annexin V 和JC-1、Tunel 细胞凋亡检测的可以应用到植物或是细菌（原核生物） 吗？

A：可以，但是需要制备原生质体，因为植物细胞或是细菌（原核生物）含有细胞壁，具体的染液使用剂量只需浸没细胞即可，染色时间对于不同细胞有一定的不同。

Q：40302ES Annexin V-FITC/PI 细胞凋亡检测试剂盒里的PI的浓度是多少呢？

A：20ug/ml。

Q:实验结果如何判断？

A：活细胞（Annexin V-/PI-）

 早期凋亡细胞（Annexin V+/PI-）

 晚期凋亡细胞和坏死细胞呈现双阳性（Annexin V+/PI+）

 裸核（Annexin V-/PI+）

Q: ​Annexin V和TUNEL有什么区别？

A：末端脱氧核苷酸转移酶 dUTP 缺口末端标记 (TUNEL) 是一种染色方法，用于识别细胞内 DNA 片段化位点——晚期细胞凋亡的标志性特征。 它使用酶末端脱氧核苷酸转移酶 (TdT) 将修饰的 dNTP（例如 dUTP）连接到片段化 DNA 链的 3'-羟基末端。 dNTPs 通常用荧光团修饰以促进量化和/或可视化。

Annexin V 染色通过结合由于细胞膜不对称性丧失而暴露在细胞外的 PS 残基来识别细胞凋亡的早期阶段。 Annexin V 通常用 FITC 等荧光团标记，以促进凋亡细胞的检测。

Q: ​1×Binding Buffer 的成分是什么？可以用什么替代？

A：成分信息保密；可以用PBS或者Hanks缓冲液代替，但是要补加2.5mM 的CaCl2。

Q: ​是否可以仅使用Annexin V-FITC进行检测而不检测PI？

A：可以的。对于特殊的检测样本，如红细胞无细胞核，如使用药物Doxorubicin（阿霉素）对PI检测存在干扰，对于这两种情况可以仅使用Annexin V-FITC进行检测。

Q: ​是否可以对组织切片进行染色（冰冻切片或石蜡切片）？

A：不可以。

[1] Du Y, Liang Z, Wang S, et al. Human pluripotent stem-cell-derived islets ameliorate diabetes in non-human primates. Nat Med. 2022;28(2):272-282. doi:10.1038/s41591-021-01645-7(IF:53.440)
[2] Chen Q, Zhang F, Dong L, et al. SIDT1-dependent absorption in the stomach mediates host uptake of dietary and orally administered microRNAs. Cell Res. 2021;31(3):247-258. doi:10.1038/s41422-020-0389-3(IF:25.617)
[3] Wang Z, Yu L, Wang Y, et al. Dynamic Adjust of Non-Radiative and Radiative Attenuation of AIE Molecules Reinforces NIR-II Imaging Mediated Photothermal Therapy and Immunotherapy. Adv Sci (Weinh). 2022;9(8):e2104793. doi:10.1002/advs.202104793(IF:16.806)
[4] Zhang M, Shao W, Yang T, et al. Conscription of Immune Cells by Light-Activatable Silencing NK-Derived Exosome (LASNEO) for Synergetic Tumor Eradication [published online ahead of print, 2022 Jun 4]. Adv Sci (Weinh). 2022;e2201135. doi:10.1002/advs.202201135(IF:16.806)
[5] Wang Z, Gong X, Li J, et al. Oxygen-Delivering Polyfluorocarbon Nanovehicles Improve Tumor Oxygenation and Potentiate Photodynamic-Mediated Antitumor Immunity. ACS Nano. 2021;15(3):5405-5419. doi:10.1021/acsnano.1c00033(IF:15.881)
[6] Li Y, Cui K, Zhang Q, et al. FBXL6 degrades phosphorylated p53 to promote tumor growth. Cell Death Differ. 2021;28(7):2112-2125. doi:10.1038/s41418-021-00739-6(IF:15.828)
[7] Li X, Yong T, Wei Z, et al. Reversing insufficient photothermal therapy-induced tumor relapse and metastasis by regulating cancer-associated fibroblasts. Nat Commun. 2022;13(1):2794. Published 2022 May 19. doi:10.1038/s41467-022-30306-7(IF:14.919)
[8] Chen YY, Ge JY, Zhu SY, Shao ZM, Yu KD. Copy number amplification of ENSA promotes the progression of triple-negative breast cancer via cholesterol biosynthesis. Nat Commun. 2022;13(1):791. Published 2022 Feb 10. doi:10.1038/s41467-022-28452-z(IF:14.919)
[9] Wang XS, Zeng JY, Li MJ, Li QR, Gao F, Zhang XZ. Highly Stable Iron Carbonyl Complex Delivery Nanosystem for Improving Cancer Therapy. ACS Nano. 2020;14(8):9848-9860. doi:10.1021/acsnano.0c02516(IF:14.588)
[10] Wang M, Zhang L, Cai Y, et al. Bioengineered Human Serum Albumin Fusion Protein as Target/Enzyme/pH Three-Stage Propulsive Drug Vehicle for Tumor Therapy [published online ahead of print, 2020 Nov 17]. ACS Nano. 2020;10.1021/acsnano.0c07610. doi:10.1021/acsnano.0c07610(IF:14.588)
[11] Deng RH, Zou MZ, Zheng D, et al. Nanoparticles from Cuttlefish Ink Inhibit Tumor Growth by Synergizing Immunotherapy and Photothermal Therapy. ACS Nano. 2019;13(8):8618-8629. doi:10.1021/acsnano.9b02993(IF:13.903)
[12] Zhao H, Xu J, Huang W, et al. Spatiotemporally Light-Activatable Platinum Nanocomplexes for Selective and Cooperative Cancer Therapy. ACS Nano. 2019;13(6):6647-6661. doi:10.1021/acsnano.9b00972(IF:13.903)
[13] Zhang C, Gao F, Wu W, et al. Enzyme-Driven Membrane-Targeted Chimeric Peptide for Enhanced Tumor Photodynamic Immunotherapy. ACS Nano. 2019;13(10):11249-11262. doi:10.1021/acsnano.9b04315(IF:13.903)
[14] Wan SS, Cheng Q, Zeng X, Zhang XZ. A Mn(III)-Sealed Metal-Organic Framework Nanosystem for Redox-Unlocked Tumor Theranostics. ACS Nano. 2019;13(6):6561-6571. doi:10.1021/acsnano.9b00300(IF:13.903)
[15] Wei JL, Wu SY, Yang YS, et al. GCH1 induces immunosuppression through metabolic reprogramming and IDO1 upregulation in triple-negative breast cancer. J Immunother Cancer. 2021;9(7):e002383. doi:10.1136/jitc-2021-002383(IF:13.751)
[16] Wang L, Qin W, Xu W, et al. Bacteria-Mediated Tumor Therapy via Photothermally-Programmed Cytolysin A Expression. Small. 2021;17(40):e2102932. doi:10.1002/smll.202102932(IF:13.281)
[17] Wan SS, Zhang L, Zhang XZ. An ATP-Regulated Ion Transport Nanosystem for Homeostatic Perturbation Therapy and Sensitizing Photodynamic Therapy by Autophagy Inhibition of Tumors. ACS Cent Sci. 2019;5(2):327-340. doi:10.1021/acscentsci.8b00822(IF:12.837)
[18] Sun D, Zou Y, Song L, et al. A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer. Acta Pharm Sin B. 2022;12(1):378-393. doi:10.1016/j.apsb.2021.06.005(IF:11.614)
[19] Yang Y, Hu D, Lu Y, et al. Tumor-targeted/reduction-triggered composite multifunctional nanoparticles for breast cancer chemo-photothermal combinational therapy. Acta Pharm Sin B. 2022;12(6):2710-2730. doi:10.1016/j.apsb.2021.08.021(IF:11.614)
[20] Hu Q, Jia L, Zhang X, Zhu A, Wang S, Xie X. Accurate construction of cell membrane biomimetic graphene nanodecoys via purposeful surface engineering to improve screening efficiency of active components of traditional Chinese medicine. Acta Pharm Sin B. 2022;12(1):394-405. doi:10.1016/j.apsb.2021.05.021(IF:11.614)
[21] Wang M, Xu Y, Zhang Y, et al. Deciphering the autophagy regulatory network via single-cell transcriptome analysis reveals a requirement for autophagy homeostasis in spermatogenesis. Theranostics. 2021;11(10):5010-5027. Published 2021 Mar 5. doi:10.7150/thno.55645(IF:11.556)
[22] Xu X, Han C, Zhang C, Yan D, Ren C, Kong L. Intelligent phototriggered nanoparticles induce a domino effect for multimodal tumor therapy. Theranostics. 2021;11(13):6477-6490. Published 2021 Apr 19. doi:10.7150/thno.55708(IF:11.556)
[23] Fan Q, Zuo J, Tian H, et al. Nanoengineering a metal-organic framework for osteosarcoma chemo-immunotherapy by modulating indoleamine-2,3-dioxygenase and myeloid-derived suppressor cells. J Exp Clin Cancer Res. 2022;41(1):162. Published 2022 May 3. doi:10.1186/s13046-022-02372-8(IF:11.161)
[24] Lei X, Cao K, Chen Y, et al. Nuclear Transglutaminase 2 interacts with topoisomerase II⍺ to promote DNA damage repair in lung cancer cells. J Exp Clin Cancer Res. 2021;40(1):224. Published 2021 Jul 5. doi:10.1186/s13046-021-02009-2(IF:11.161)
[25] Xu L, Wang Y, Song E, Song Y. Nucleophilic and redox properties of polybrominated diphenyl ether derived-quinone/hydroquinone metabolites are responsible for their neurotoxicity. J Hazard Mater. 2021;420:126697. doi:10.1016/j.jhazmat.2021.126697(IF:10.588)
[26] Zhang C, Peng SY, Hong S, et al. Biomimetic carbon monoxide nanogenerator ameliorates streptozotocin induced type 1 diabetes in mice. Biomaterials. 2020;245:119986. doi:10.1016/j.biomaterials.2020.119986(IF:10.317)
[27] Zhang L, Cheng Q, Li C, Zeng X, Zhang XZ. Near infrared light-triggered metal ion and photodynamic therapy based on AgNPs/porphyrinic MOFs for tumors and pathogens elimination. Biomaterials. 2020;248:120029. doi:10.1016/j.biomaterials.2020.120029(IF:10.317)
[28] Zhang C, Zheng DW, Li CX, et al. Hydrogen gas improves photothermal therapy of tumor and restrains the relapse of distant dormant tumor. Biomaterials. 2019;223:119472. doi:10.1016/j.biomaterials.2019.119472(IF:10.273)
[29] Cheng Q, Yu W, Ye J, et al. Nanotherapeutics interfere with cellular redox homeostasis for highly improved photodynamic therapy. Biomaterials. 2019;224:119500. doi:10.1016/j.biomaterials.2019.119500(IF:10.273)