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腺相关病毒(AAV)滴度快速检测试剂盒

发布者:艾美捷科技    发布时间:2021-08-11     
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【前情提示】慢病毒滴度检测

 

腺相关病毒(adeno-associatedvirus,AAV),也称腺伴随病毒,属于微小病毒科依赖病毒属,是目前发现的一类结构最简单的单链DNA缺陷型病毒,需要辅助病毒(通常为腺病毒)参与复制。AAV基因组为4.8kb的线状单链DNA;AAV免疫原性低,基因持续表达半年以上;AAV具有多种血清型,可以特异性靶向感染不同的组织或器官,是动物活体基因转导的首选工具!

 

腺相关病毒.jpg

 

腺相关病毒血清型众多(12种),不同血清型对不同组织亲和性不同。如何选择呢?(如下图)

 

腺相关病毒血清型.png

 

通过病毒载体传递基因的一个特殊挑战是病毒滴度的准确测量。传统上,AAV颗粒是通过DNA斑点印迹或类似方法测量的。这些方法非常耗时,并且存在高度的测定间变异性。对于高度纯化的病毒样本,260nm处的吸光度已用于估计病毒颗粒的总数。然而,这种方法不能用于未纯化的病毒上清液,因为它包含的其他成分会导致260nm的吸光度。通过使用仅与AAV完整颗粒反应的抗体开发了一种ELISA方法;然而,这种方法测量所有AAV颗粒,包括那些缺乏基因组DNA的颗粒。

 

那么如何真正准确可靠地检测腺相关病毒(AAV)滴度呢?作为专业的生命科学医药原料供应商,艾美捷科技为您推荐:众多高分文章引用的,腺相关病毒(AAV)滴度检测试剂盒(适用于任何血清型的纯化AAV,以及未纯化的AAV-2和AAV-DJ型)

 

产品腺相关病毒(AAV)滴度检测试剂盒
QuickTiter AAV Quantitation Kit
货号CBL-VPK-145
检测方法荧光法
适用病毒类型任何血清型的纯化AAV,以及未纯化的AAV-2和AAV-DJ型
说明书下载点击下载
实验优势1.简单快捷:未纯化的上清液大约需要 4 小时,对于纯化的 AAV 大约需要 30 分钟;
2.样本适用:检测任何血清型的纯化AAV,以及未纯化的AAV-2和AAV-DJ型
3.灵敏度:可检测1X109GC/ml的AAV的细胞上清;检测5X1010GC/ml的纯化的AAV。
试剂盒组分1.ViraBind AAV Reagent A
2.ViraBind AAV Reagent B
3.QuickTiter AAV Capture Matrix
4.QuickTiter AAV Wash Solution (5X)
5.QuickTiter Solution C (10X)
6.CyQuant GR Dye (400X)
7.QuickTiter AAV DNA Standard
保存条件收到后,在室温下保存ViraBind AAV试剂B,其它所有试剂组分保存在4℃。

* 本产品仅适用于科研用途.

 

腺相关病毒(AAV)滴度检测试剂盒原理与标曲展示:

腺相关病毒(AAV)滴度检测试剂盒原理.jpg

腺相关病毒(AAV)滴度检测试剂盒标曲.jpg

腺相关病毒(AAV)滴度检测试剂盒原理腺相关病毒(AAV)滴度检测试剂盒标曲

 

腺相关病毒(AAV)滴度检测试剂盒的Nature结果鉴赏:

腺相关病毒(AAV)滴度检测试剂盒的Nature结果鉴赏.jpg

Nat-Neurosci-1.jpg

Nat Neurosci. doi: 10.1038/s41593-020-0674-y.

 

近期发表文章:

1.Francisco, J. et al. (2021). AAV-mediated YAP expression in cardiac fibroblasts promotes inflammation and increases fibrosis. Sci Rep. 11(1):10553. doi: 10.1038/s41598-021-89989-5.

2.Kwon, O.C. et al. (2021). SGK1 inhibition in glia ameliorates pathologies and symptoms in Parkinson disease animal models. EMBO Mol Med. doi: 10.15252/emmm.202013076.

3.Lebeau, P.F. et al. (2020). The loss-of-function PCSK9Q152H variant increases ER chaperones GRP78 and GRP94 and protects against liver injury. J Clin Invest. doi: 10.1172/JCI128650.

4.Kawaguchi, Y. et al. (2020). Endoplasmic reticulum chaperone BiP/GRP78 knockdown leads to autophagy and cell death of arginine vasopressin neurons in mice. Sci Rep. 10(1):19730. doi: 10.1038/s41598-020-76839-z.

5.Rai, R. et al. (2020). Targeted gene correction of human hematopoietic stem cells for the treatment of Wiskott - Aldrich Syndrome. Nat Commun. 11(1):4034. doi: 10.1038/s41467-020-17626-2.

6.Zeng, J. et al (2020). The Zika Virus Capsid Disrupts Corticogenesis by Suppressing Dicer Activity and miRNA Biogenesis. Cell Stem Cell. S1934-5909(20)30350-7. doi: 10.1016/j.stem.2020.07.012.

7.Tang, Y. et al. (2020). Social touch promotes interfemale communication via activation of parvocellular oxytocin neurons. Nat Neurosci.doi: 10.1038/s41593-020-0674-y.

8.Zhu, J. et al. (2020). Preparation of a Bacteriophage T4-based Prokaryotic-eukaryotic Hybrid Viral Vector for Delivery of Large Cargos of Genes and Proteins into Human Cells. Bio-protocol. 10(07): e3573. doi: 10.21769/BioProtoc.3573.

9.Wu, Z. et al. (2020). Gene therapy conversion of striatal astrocytes into GABAergic neurons in mouse models of Huntington's disease. Nat Commun. 11(1):1105. doi: 10.1038/s41467-020-14855-3.

10.Zhang, H. et al. (2020). Vitamin D receptor targets hepatocyte nuclear factor 4α and mediates protective effects of vitamin D in nonalcoholic fatty liver disease. J Biol Chem. pii: jbc.RA119.011487. doi: 10.1074/jbc.RA119.011487.

11.Xu, Y. et al. (2020). Diabetic nephropathy execrates epithelial-to-mesenchymal transition (EMT) via miR-2467-3p/Twist1 pathway. Biomed Pharmacother. 125:109920. doi: 10.1016/j.biopha.2020.109920.

12.Oser, M.G. et al. (2019). The KDM5A/RBP2 histone demethylase represses NOTCH signaling to sustain neuroendocrine differentiation and promote small cell lung cancer tumorigenesis. Genes Dev. doi: 10.1101/gad.328336.119.

13.Niranjan, N. et al. (2019). Sarcolipin overexpression impairs myogenic differentiation in Duchenne muscular dystrophy. Am J Physiol Cell Physiol. doi: 10.1152/ajpcell.00146.2019.

14.Ferretti, V. et al. (2019). Oxytocin Signaling in the Central Amygdala Modulates Emotion Discrimination in Mice. Curr Biol. pii: S0960-9822(19)30499-3. doi: 10.1016/j.cub.2019.04.070.

15.Hasan, M.T. et al. (2019). A Fear Memory Engram and Its Plasticity in the Hypothalamic Oxytocin System. Neuron. pii: S0896-6273(19)30386-1. doi: 10.1016/j.neuron.2019.04.029.

16.Lee, S. et al. (2019). Anti-EpCAM-conjugated adeno-associated virus serotype 2 for systemic delivery of EGFR shRNA: Its retargeting and antitumor effects on OVCAR3 ovarian cancer in vivo. Acta Biomater. pii: S1742-7061(19)30287-9. doi: 10.1016/j.actbio.2019.04.044.

17.Nakamura, M. et al. (2019). Glycogen Synthase Kinase-3α Promotes Fatty Acid Uptake and Lipotoxic Cardiomyopathy. Cell Metab. pii: S1550-4131(19)30005-1. doi: 10.1016/j.cmet.2019.01.005.

18.Tseng, S.J. et al. (2018). Targeting Tumor Microenvironment by Bioreduction-Activated Nanoparticles for Light-Triggered Virotherapy. ACS Nano. 12(10):9894-9902. doi: 10.1021/acsnano.8b02813.

19.Ikegami, R. et al. (2018). Gamma-Aminobutyric Acid Signaling in Brown Adipose Tissue Promotes Systemic Metabolic Derangement in Obesity. Cell Rep. 24(11):2827-2837.e5. doi: 10.1016/j.celrep.2018.08.024.

20.Menon, R. et al. (2018). Oxytocin Signaling in the Lateral Septum Prevents Social Fear during Lactation. Curr Biol. 28(7):1066-1078.e6. doi: 10.1016/j.cub.2018.02.044.

21.Wen, L. et al. (2018). Transient High Pressure in Pancreatic Ducts Promotes Inflammation and Alters Tight Junctions via Calcineurin Signaling in Mice. Gastroenterology. 155(4):1250-1263.e5. doi: 10.1053/j.gastro.2018.06.036.

22.McGrady NR, et al. (2017). Upregulation of the endothelin A (ETA) receptor and its association with neurodegeneration in a rodent model of glaucoma. BMC Neurosci. 18(1):27. doi: 10.1186/s12868-017-0346-3.

23.Wang, P., et al. (2017). Tau interactome mapping based identification of Otub1 as Tau deubiquitinase involved in accumulation of pathological Tau forms in vitro and in vivo. Acta Neuropathol. doi:10.1007/s00401-016-1663-9.

24.Tseng, S.J. et al. (2016). Remote control of light-triggered virotherapy. ACS Nano 10:10339-10346.

25.Eliava, M. et al. (2016). A new population of parvocellular oxytocin neurons controlling magnocellular neuron activity and inflammatory pain processing. Neuron 89:1291-1304.

26.Madro al, N. et al. (2016). Rapid erasure of hippocampal memory following inhibition of dentate gyrus granule cells. Nat Commun. doi:10.1038/ncomms10923.

27.Park, H. Y. et al. (2016). Gadd45β ameliorates L-DOPA-induced dyskinesia in a Parkinson's disease mouse model. Neurobiol Dis. 89:169-179.

28.Tran, L. & Keele, N. B. (2016). CaMKIIα knockdown decreases anxiety in the open field and low serotonin-induced upregulation of GluA1 inthe basolateral amygdala. Behav Brain Res. 303:152-159.

29.Chen, W. et al. (2016). The silencing of cathepsin K used in gene therapy for periodontal disease reveals the role of cathepsin K in chronic infection and inflammation. J Periodontal Res.doi:10.1111/jre.12345.

30.Orabi, A. I. et al. (2015). Dynamic imaging of pancreatic NF-κB activation in live mice using AAV infusion and ioluminescence. J Biol Chem. doi:10.1074/jbc.M115.647933

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