计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-80℃储存。特低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
AAV-mCherry-GFP-LC3B ，即AAV9 serotype adeno-associated virus expressing mCherry-GFP-LC3B fusion protein，是一种可表达mCherry-GFP-LC3B融合蛋白的腺相关病毒，用于注射小鼠等动物后进行细胞自噬(autophagy)检测。LC3是酵母自噬关键蛋白ATG8在哺乳动物中的同源蛋白。LC3最初被分析鉴定为microtubule-associated protein 1 light chain 3 (MAP1LC3)。LC3蛋白家族包括LC3A、B、C和GABARAP等亚家族，其中对于LC3B的研究最为广泛。到目前为止，LC3B被认为是细胞自噬信号通路最为关键的标志性蛋白。LC3B是一个具有125个氨基酸残基的蛋白，在蛋白合成后，被Atg4所酶切而失去了C端的22个氨基酸蛋白，从而暴露出C端的甘氨酸，人们把它命名为胞质形式的LC3B I。在细胞自噬的过程中，其C端暴露的甘氨酸经过一个类似泛素化的过程，以ATG7为E1样激活酶，以ATG3为E2样连接酶，以Atg12-Atg5-Atg16复合物为E3样连接酶，在C端甘氨酸上共价连接上磷脂酰乙醇胺(phosphatidylethanolamine, PE)而成为LC3B-PE即LC3B II。与胞质定位的LC3B I不同，LC3B II定位于自噬体(autophagosome)的内膜和外膜上。在自噬体与溶酶体融合后，自噬体外膜上的LC3B II被Atg4所酶切，而自噬体内膜上的LC3B II则被溶酶体内的蛋白酶所降解。尽管LC3B II比LC3B I的分子量要大，但由于其极强的疏水性，在SDS-PAGE电泳时，LC3B II比LC3B I迁移得更快，其表观分子量分别为14kD和16kD。自噬(Autophagy)是一种在进化上高度保守的通过溶酶体吞噬并降解部分自身组分的细胞内分解代谢途径。自噬与多种生理功能有关，在饥饿等不利的环境条件下，细胞通过自噬降解多余或异常的细胞内组分，为细胞的生存提供能量及原材料，促进生物体的生长发育、细胞分化及对环境变化产生应答。自噬异常与多种病理过程如肿瘤、神经退行性疾病、代谢疾病、病原体感染等都有密切关系。由于细胞自噬在生理和病理过程中都有重要作用，自噬已经成为细胞生物学领域的一个新的研究热点。AAV-mCherry-GFP-LC3B是阿拉丁自行研发的重组腺相关病毒，感染后能够在靶细胞中有效表达红色荧光蛋白mCherry、绿色荧光蛋白(GFP)和LC3B的融合蛋白，呈现明亮的红色和绿色荧光，可以用于细胞自噬的检测。自噬小体在与溶酶体融合过程中，溶酶体内的酸性环境会导致GFP荧光淬灭，这为追踪GFP-LC3B的细胞定位增加了难度。mCherry是一种来自于蘑菇珊瑚(mushroom coral)的单体红色荧光蛋白，当其与GFP进行LC3B的共同标记时，在GFP被溶酶体酸性环境所淬灭的情况下，可以发出红色荧光的mCherry因为其卓越的稳定性而被保留。因此，通过融合表达mCherry-GFP-LC3B蛋白，可以非常有效地追踪自噬过程。在用AAV-mCherry-GFP-LC3B腺相关病毒感染细胞后，在非自噬的情况下，荧光显微镜下mCherry-GFP-LC3B以弥散的黄色荧光(mCherry和GFP的综合效果)形式存在于细胞质中；而在自噬的情况下，荧光显微镜下mCherry-GFP-LC3B则聚集在自噬体膜上，以黄色斑点的形式表现出来(LC3B dot or punctae)；当自噬体与溶酶体融合后，因GFP荧光的部分淬灭而以红色斑点的形式表现出来。腺相关病毒感染细胞后通常不会与基因组DNA重组，但可以长期稳定地表达目的蛋白，特别是可以在很多不分裂的细胞中长期稳定表达。AAV还具有滴度高，体内注射不产生明显的免疫反应，对于增殖和非增殖细胞感染效率高等优点。阿拉丁的AAV-mCherry-GFP-LC3B是复制缺陷型的，感染普通的细胞后不能进行复制和扩增，必须在辅助病毒存在的情况下才能复制和扩增，从而有效降低了本产品在活体生物中的风险。腺相关病毒目前发现主要有12种血清型，不同血清型AAV有不同的衣壳蛋白，其识别结合的细胞膜表面受体也有所不同，因此不同血清型AAV感染不同类型细胞的感染效率也各不相同。不同血清型AAV的组织亲和性参见下表。本产品为组织亲和性最为广泛的AAV9血清型，能感染多种组织，使用相对比较广泛。不同血清型AAV的感染不同种类体外培养细胞的效率参见下表(以AAV2感染效率100%为标准)。参考下表，本产品感染培养细胞的效率很低，不太适合用于感染培养的细胞。实验室常用的逆转录病毒、慢病毒、腺相关病毒和腺病毒的主要特之间的比较和差别参见下表。具体的特定病毒的一些特征和下表相比可能会有一定差异。阿拉丁提供腺病毒和腺相关病毒两种类型的表达mCherry-GFP-LC3B的产品。两种产品的比较和选择请参见下表。其中C3013 AAV-mCherry-GFP-LC3B适合用于动物实验， C3011 AD-mCherry-GFP-LC3B更适合于细胞实验，由于表达起始时间短，适合用于能被腺病毒感染并且希望快速获得实验结果的情况。另外，C3011 AD-mCherry-GFP-LC3B可以在表达E1的HEK293A、HEK293等适当细胞中扩增，便于后续长期使用。本产品的滴度为1×1013vp/ml。vp, viral particle，即病毒颗粒。如果用于尾静脉注射8-12周的小鼠，每只小鼠的推荐注射量为2-5×1011vp，此时一个包装的本产品可以用于注射5-12只小鼠。用于小鼠脑内(intracerebal)和心室内(intraventricular)注射的推荐用量是1-5×109vp，用于鼻内(intranasal)注射时的推荐用量为1-3×1010vp，用于眼内(intravitreal)注射时的推荐用量为1-4×109vp。
注意事项：
反复冻融会降低病毒滴度，如有必要请在收到本产品后分装保存。分装时必须在冰浴上进行。病毒融解后，如果在一周内使用，可以放置于4℃。如果-80℃保存时间超过一年，可能会导致滴度下降，此时建议重新测定病毒滴度。本产品对于小鼠等动物在体感染的效率很高，但对大多数常见的体外培养细胞的感染效率很低。因此，本产品适合用于小鼠等动物的体内感染，不太适合用于培养细胞的感染。如果用于培养细胞的自噬研究，推荐选购C3011 AD-mCherry-GFP-LC3B。如果希望使用本产品之外的其它血清型的AAV-mCherry-GFP-LC3B，可以联系阿拉丁的技术服务部进行定制。对于一些比较珍贵的基因敲除或转基因动物或者疾病模型动物，建议先注射1-2个野生型正常小鼠进行预实验，确定注射方式和注射剂量后再开始正式实验。本产品使用前请仔细阅读附录1《腺相关病毒使用安全规范》。本产品生物安全等级为Biosafety Level 1 (BSL-1)，没有确凿证据显示会导致健康成人产生疾病(Not known to consistently cause diseases in healthy adults)，可以按照常规的微生物实验操作要求进行操作。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。
使用说明：
1.根据拟感染的动物组织，选择合适的特定血清型的AAV。AAV9血清型使用最为广泛，可以感染多种小鼠组织，具体请参见本说明书的不同血清型AAV的组织亲和性表。 2.根据拟感染的动物组织，通过尾静脉注射、局部注射等方式注射本产品。如果用于尾静脉注射8-12周的小鼠，每只小鼠的推荐注射量为2-5×1011vp；用于小鼠脑内(intracerebal)和心室内(intraventricular)注射的推荐用量是1-5×109vp；用于鼻内(intranasal)注射时的推荐用量为1-3×1010vp；用于眼内(intravitreal)注射时的推荐用量为1-4×109vp。对于特定组织自噬研究的最佳注射剂量，请参考文献数据或自行通过预实验进行摸索。 3.通常注射后3-4天即可以检测到mCherry-GFP-LC3B融合蛋白的表达，也可以在注射后更长时间检测mCherry-GFP-LC3B融合蛋白的表达，以确定特定组织中的细胞自噬情况。附录： 1.腺相关病毒使用安全规范a.作为一种相对安全的病毒，尽管腺相关病毒基因组在感染后通常不会整合进入宿主细胞基因组中，也不会在细胞体内进行复制，但是仍然具有可能的潜在生物学危险。我们建议使用者在病毒操作前应仔细阅读本规范，并在实验中严格按照本规范的要求进行操作。更为严格的美国CDC的生物安全等级及其操作与防护要求参考附表1，也可以访问如下网页：https://www.cdc.gov/labs/pdf/CDC-BiosafetyMicrobiologicalBiomedicalLaboratories-2020-P.pdf。b.腺相关病毒操作时应使用相应级别的生物安全柜，不同的腺相关病毒的生物安全等级会有所不同。如果使用普通超净工作台操作病毒，请不要打开排风机，以尽量避免可能污染病毒的尘埃正面吹向操作人员而被吸入。c.实验操作时必需佩戴一次性帽子、口罩，穿戴实验手套及专门的实验服，避免身体直接接触病毒。手部及面部有开放性创口时，禁止进行病毒操作。d.操作病毒时需小心谨慎，不要产生气雾或飞溅。如果操作时超净工作台或其它器皿上有病毒污染，请立即用70%乙醇或2%SDS溶液擦拭干净，或者采取其它的妥善措施。e.如果需要离心，应使用密封性好的离心管，或用封口膜密封后离心，最好使用专门的离心机。f.用显微镜观察细胞感染情况时应遵从以下步骤：拧紧培养瓶或盖紧培养板，用70%乙醇清理培养瓶或培养板外壁后到显微镜处观察拍照。离开显微镜实验台之前，用70%乙醇擦洗显微镜实验台。g.所有被病毒污染过的枪头、离心管、培养板(皿、瓶)、培养液、手套等，在丢弃前请用84消毒液或2%SDS浸泡过夜。脱掉手套后，用肥皂或洗手液清洗双手。h.病毒飞溅或是含有病毒的气溶胶与人体接触，须用大量清水冲洗眼睛、皮肤或粘膜等接触的部位至少15min。i.含病毒的针头或是其它利器刺破皮肤，伤口应立即用10%的碘伏溶液擦洗数分钟，然后用大量清水冲洗。2.病毒MOI的计算MOI (Multiplicity of Infection)定义：病毒感染细胞时，病毒数量与细胞数量的比值。Pfu (Plaque forming units)定义：具有生物活性的病毒颗粒数量。可以根据如下公式计算所需pfu：所需pfu=细胞数量×MOI例如，需要向1×105细胞中加入2 MOI的病毒，即所需pfu=(1×105 cells)×(2 MOI)=2×105 pfu。若病毒母液滴度为1×108 pfu/ml，则细胞培养液中应加入(2×105 pfu)/(1×108 pfu/ml)=0.002ml病毒母液，即2μl病毒母液。附表1. 生物安全等级及其操作与防护要求Table 1. Summary of Recommended Biosafety Levels for Infectious AgentsBSLAgentsPracticesPrimary Barriers and Safety EquipmentFacilities (Secondary Barriers)1Not known to consistently cause diseases in healthy adultsStandard microbiological practices■ No primary barriers required.■ PPE: laboratory coats and gloves; eye, face protection, as neededLaboratory bench and sink required2■ Agents associated with human disease■ Routes of transmission include perxadcutaneous injury, ingestion, mucous membrane exposureBSL-1 practice plus:■ Limited access■ Biohazard warning signs■ “Sharps” precautions■ Biosafety manual defining any needed waste decontamination or medical surveillance policiesPrimary barriers:■ BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials■ PPE: Laboratory coats, gloves, face and eye protection, as neededBSL-1 plus:■ Autoclave available3Indigenous or exotic agents that may cause serious or potentially lethal disease through the inhalation route of exposureBSL-2 practice plus:■ Controlled access■ Decontamination of all wastePrimary barriers:■ BSCs or other physical containment devices used for all open manipulaxadtions of agents■ PPE: Protective laboratory clothing, gloves, face, eye and respiratory protection, as neededBSL-2 plus:■ Physical separation from access corridors■ Self-closing, double-door access■ Negative airflow into laboratory■ Entry through airlock or anteroom■ Hand washing sink near laboratory exit4■ Dangerous/exotic agents which post high individual risk of aerosol-transxadmitted laboratory infections that are frequently fatal, for which there are no vaccines or treatments■ Agents with a close or identical antixadgenic relationship to an agent requirxading BSL-4 until data are available to redesignate the level■ Related agents with unknown risk of transmissionBSL-3 practices plus:■ Clothing change before entering■ Shower on exit■ All material decontaminated on exit from facilityPrimary barriers:■ All procedures conducted in Class III BSCs or Class I or II BSCs in comxadbination with full-body, air-supplied, positive pressure suitBSL-3 plus:■ Separate building or isolated zone■ Dedicated supply and exhaust, vacuum, and decontamination systems■ Other requirements outlined in the textBSL, biosafety level; PPE, personal protective equipment.
AAV-mCherry-GFP-LC3B is an AAV9 serotype adeno-associated virus that can express mCherry-GFP-LC3B fusion protein in infected cells or tissues. It can be used for autophagy assays after injection into animals such as mice.Autophagy is a highly conserved intracellular process that degrades long-lived proteins, organelles, and other cytoplasmic components via the lysosomal pathway. Under unfavorable conditions such as starvation, cells degrade excess or abnormal intracellular components through autophagy to provide energy and raw materials for cell survival, promoting growth and development of organisms, cell differentiation, and responses to environmental changes. Abnormal autophagy is closely related to a variety of pathological processes such as tumors, neurodegenerative diseases, metabolic diseases, and pathogen infections. Because of the importance of autophagy in both physiological and pathological processes, it has become a new hot research topic in the field of cell biology.LC3 is the mammalian homolog of ATG8 which is a key protein in yeast autophagy. The LC3 protein family includes LC3A, LC3B, LC3C and GABARAP subfamilies, among which LC3B is the most extensively studied. LC3B has been considered to be the most critical marker protein in the autophagy signaling pathway. The pro-form of LC3B consisting of 125 amino acids is cleaved by Atg4 into the LC3B-I form with 22 amino acids lost at the C-terminus, thus exposing a C-terminal glycine. In the process of autophagy, LC3B-1 undergoes a ubiquitination-like process and becomes the LC3B-II form with phosphatidylethanolamine (PE) covalently linked with the C-terminal glycine, during which ATG7 acts as the E1-like activating enzyme, ATG3 as the E2-like conjugation enzyme, and the Atg12-Atg5-Atg16 complex as the E3-like ligase. Unlike LC3B-I which is localized in cytoplasm, LC3B-II is localized on the inner and the outer membranes of autophagosome. After autophagosome-lysosome fusion, LC3B-II on the outer membrane of autophagosome is cleaved by Atg4, while LC3B-II on the inner membrane is degraded by proteases in lysosome. Although LC3B-II has a larger molecular weight than LC3B-I, it migrates faster than LC3B-I during SDS-PAGE electrophoresis due to its extreme hydrophobicity, appearing 14kD and 16kD, respectively.AAV-mCherry-GFP-LC3B is a recombinant adeno-associated virus developed by aladdin, which can effectively express LC3B proteins fused with red fluorescent mCherry as well as green fluorescent GFP in target cells.In the process of autophagosome-lysosome fusion, the fluorescence of GFP will be quenched due to the acidic environment within lysosome, making it difficult to track the intracellular location of GFP-LC3B. However, mCherry is a monomeric fluorescent protein from mushroom coral and its red fluorescence is stable under acidic conditions. Therefore, the mCherry-EGFP-LC3B fusion protein enables effective tracking of autophagy processes. After infecting cells with the Ad-mCherry-GFP-LC3B adenovirus, non-autophagy cells exhibits diffusive yellow fluorescence in cytoplasm due to the combination of green and yellow fluorescence, while in cells with autophagy, mCherry-GFP-LC3B aggregates on the autophagosome membrane and appears as yellow spots (LC3B dot or punctae) or as red spots due to partial quenching of GFP fluorescence during the process of autophagosome-lysosome fusion.AAV usually does not integrate the target gene into genomic DNA of infected cells, but can stably express target proteins over time, especially in non-dividing cells. AAV also has the advantages of high titer, no significant immune response to in vivo injection, and high efficiency in infecting proliferating and non-proliferating cells.AAV-mCherry-GFP-LC3B is replication-deficient, which can only replicate and amplify in the presence of auxiliary viruses, thus effectively reducing the risk of this product in living organisms.There are mainly 12 serotypes of AAV . Different serotypes of AAV have different coat proteins recognized by different receptors on the cell membrane of infected cells. The tissue affinities of different serotypes of AAV are listed below.AAV SerotypeTissue TropismLiverMuscleBrainRetinaLungHeartPancreasKidneyAAV1√Neuron & glia√√√AAV2√√√AAV3√√√AAV4√√AAV5Neuron & glia√alveolar cellsAAV6√√√AAV7√Neuron√AAV8√√Neuron√√AAV9√√Neuron√√√√√This product is the most widely used AAV9 serotype that can infect a variety of tissues.The transduction efficiency of different cell lines in vitro by different AAV serotypes is listed below . As shown in the table, this product has a very low efficiency in transducting cultured cells and is not suitable for infecting cultured cells. Cell lineAAV1AAV2AAV3AAV4AAV5AAV6AAV8AAV9Huh-7131002.500.1100.70HEK293251002.50.10.150.70.1Hela310020.16.710.20.1HepG2310016.70.31.750.3NDHep1A201000.210.110.2091117100110.20.1170.1NDCHO100100141.433350101COS33100333.351420.5MeWo10100200.36.71010.2NIH3T3101002.92.90.3100.3NDA5491410020ND0.5100.50.1HT118020100100.10.3330.50.1Monocytes1111100NDND1251429NDNDImmature DC2500100NDND2222857NDNDMature DC2222100NDND3333333NDNDPlease refer to the table below for the comparison and the differences in the main features of retroviruses, lentiviruses, adeno-associated viruses and adenoviruses commonly used in the laboratory. The characteristics of some particular viruses may differ from those in the table below.RetrovirusLentivirusAAVAdenovirusGenomessRNA(+)ssRNA(+)ssDNAdsDNACoatEnvelopedEnvelopedNakedNakedParticle size90-100nm90-100nm20-30nm60-90nmGenome size7-10kb9kb5kb38-39kbGenome integrationYesYesNoNoPackaging capacity2.5-5kb2.5-6kb2.5-4.5kb3-8kbInfection tropismDividing cellsDividing and non-dividing cellsDividing and non-dividing cellsDividing and non-dividing cellsRelative Transduction EfficiencyND0.70.71Expression started48-72h48-72h72-96h24-48hExpression duration＞ 2 months＞ 2 months＞ 6 months3-4 weeksExpression levelMediumMediumMediumHighImmune responseLowLowVery lowHighIn vivo safetyMediumMediumHighLowTiter before concentration (IFU/ml)1061071011107Titer after concentration (IFU/ml)ND1080.5-1x10131010Able to obtain high MOINo (≤ 10 copies integrated)No (≤ 10 copies integrated)YesYesBiosafety levelBSL-2BSL-2BSL-1BSL-2aladdin offers adenovirus and adeno-associated virus for the expression of mCherry-GFP-LC3B fusion proteins in infected cells
Precautions：
Repeated freeze-thaws will reduce the viral titer. If necessary, please store in aliquots at the first use. Aliquoting must be performed on ice. This product can be stored at 4℃ if it is used within a week, but it should be noted that the viral titer decreases over the time of storage at 4℃. If stored at -80℃ for more than one year, the titer may decrease, and it is recommended to re-titrate this product before use.This product is highly efficient and suitable for in vivo infection of animals such as mice, but is inefficient and less suitable for in vitro infection of cultured cells. For autophagy studies of cultured cells, we recommend using the Ad-mCherry-GFP-LC3B (, C3011).For other serotypes of AAV-mCherry-GFP-LC3B, please contact CRO Department for customization.For some valuable knockout or transgenic animals or model animals with disease, it is recommended to perform preliminary tests with 1-2 wild type mice to determine the optimal injection method and dosage before starting formal experiments with the valuable animals.Please read Appendix 1 "Safety Specifications for Lentivirus Use" carefully before using this product. The biosafety level of this product is Biosafety Level 1 (BSL-1). It is not known to consistently cause diseases in healthy adults and can be handled with standard microbiological practices.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.
Instructions for Use：
1.Select an appropriate AAV serotype for the animal tissue to be infected. The AAV9 serotype is the most widely used and can infect a variety of mouse tissues. Please refer to the tissue affinity table for different serotypes of AAV in this manual. 2.Inject this product into animals by tail vein injection, local injection or other injection methods. The recommended dosage per 8-12 weeks mouse for tail vein injection is 2-5×1011 vp, for intracerebral and intraventricular injections is 1-5×109 vp, for intranasal injections is 1-3×1010 vp, for intravitreal injections is 1-4×109 vp. For the optimal injection dosage for autophagy studies of particular tissues, please refer to relevant literature or optimize through preliminary experiments.3.Expression of mCherry-GFP-LC3B fusion protein can usually be visible 3-4 days after injection. Observation of mCherry-GFP-LC3B fusion proteins can also be performed after a longer period of time to determine the cell autophagy in specific tissues.Appendix:1.Safety specifications for handling adeno-associated virusesa.As a relatively safe virus, although the adenovirus genome does not integrate into the host cell genome or replicate in cells after infection, it still has potential biological risks. We recommend that users read the specifications carefully before virus manipulation and operate in strict accordance with the requirements. For more stringent US CDC biosafety levels and their operation and protection requirements, please refer to Appendix 1 or visit the following website: http://www.cdc.gov/biosafety/publications/bmbl5/BMBL5_sect_IV.pdf.b.Biosafety cabinets of corresponding levels should be used for adenovirus operations, and the biosafety levels vary for different adenoviruses. When using an ordinary laminar flow hood to operate the virus, please do not turn on the exhaust fan to avoid the dust that may be virus-contaminated being blown to the operator and inhaled.c.Disposable hats, masks, laboratory gloves and special laboratory coats must be worn during the experiments to avoid direct contact with the virus. Virus manipulation is prohibited when there are open wounds on the hands and face.d.Be careful not to produce aerosol or splash when handling the virus. If the the laminar flow hood or other utensils are contaminated by the virus during operation, please wipe them with 70% ethanol or 2% SDS solution immediately, or take other appropriate measures.e.If centrifugation is required, use a well-sealed centrifuge tube, or seal it with parafilm and centrifuge, preferably using a centrifuge designated for virus operation.f.The following steps should be followed when observing cell infection under a microscope: Tighten the culture flask or cover the culture plate, clean the outer surface of the culture flask or culture plate with 70% ethanol, and then examine by microscope. After finishing examination, clean the microscope bench with 70% ethanol.g.All virus-contaminated pipette tips, centrifuge tubes, culture plates (dishes, bottles), culture solution, gloves, and other consumables should be soaked in disinfectant or 2% SDS overnight before discarding.h.After removing gloves, wash hands with soap or hand sanitizer.i.When the virus is splashed or the aerosol containing the virus comes into contact with human body, flush the eyes, skin or mucous membranes with plenty of water for at least 15 minutes.j.When the skin is punctured by a needle or other sharp object containing the virus, the wound should be immediately sterilized with 10% iodine solution for several minutes and then rinsed with plenty of water.2.Calculation of viral MOIMOI (Multiplicity of Infection) is defined as the ratio of the number of viruses to the number of cells when virus infects cells.Pfu (plaque forming units) defines the number of biologically active virus particles.The required pfu can be calculated according to the following formula.Required pfu = number of cells x MOIFor example: If 2 MOI of virus is needed for infecting 1 × 105 cells, the required pfu = (1 × 105 cells) × (2 MOI) = 2 × 105 pfu. For a virus stock with a titer of 1×108 pfu/ml, the volume of the virus stock required will be 2×105 pfu / (1×108 pfu/ml) × 1000 = 2μl.Table 1. Biosafety levels and their handling and protection requirementsBSLAgentsPracticesPrimary Barriers andSafety EquipmentFacilities(Secondary Barriers)1Not known to consistently cause diseases in healthy adultsStandard microbiological practices■ No primary barriers required.■ PPE: laboratory coats and gloves; eye, face protection, as neededLaboratory bench and sink required2■ Agents associated with human disease■ Routes of transmission include percutaneous injury, ingestion, mucous membrane exposureBSL-1 practice plus: ■ Limited access■ Biohazard warning signs■ “Sharps” precautions■ Biosafety manual defining any needed waste decontamination or medical surveillance policiesPrimary barriers:■ BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials■ PPE: Laboratory coats, gloves, face and eye protection, as neededBSL-1 plus:■ Autoclave available3Indigenous or exotic agents that may cause serious or potentially lethal disease through the inhalation route of exposureBSL-2 practice plus: ■ Controlled access■ Decontamination of all waste■ Decontamination of laboratory clothing before launderingPrimary barriers:■ BSCs or other physical containment devices used for all open manipulations of agents■ PPE: Protective laboratory clothing, gloves, face, eye and respiratory protection, as neededBSL-2 plus:■ Physical separation from access corridors■ Self-closing, double-door access■ Exhausted air not recirculated■ Negative airflow into laboratory■ Entry through airlock or anteroom■ Hand washing sink near laboratory exit4■ Dangerous/exotic agents which post high individual risk of aerosol-transmitted laboratory infections that are frequently fatal, for which there are no vaccines or treatments■ Agents with a close or identical antigenic relationship to an agent requiring BSL-4 until data are available to redesignate the level■ Related agents with unknown risk of transmissionBSL-3 practices plus:■ Clothing change before entering■ Shower on exit■ All material decontaminated on exit from facilityPrimary barriers:■ All procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive pressure suitBSL-3 plus:■ Separate building or isolated zone■ Dedicated supply and exhaust, vacuum, and decontamination systems■ Other requirements outlined in the textBSL, biosafety level; PPE, personal protective equipment.
规格
产品名称
AAV-mCherry-GFP-LC3B(AAV9)
规格或纯度
10^13vp/ml
分子类型
基因
产品线
生物产品
储存与运输
浓度
10^13vp/ml
储存条件
-80℃储存
运输条件
特低温运输
稳定性与储存
-80℃保存，一年有效。-20℃保存，1-2个月内有效。4℃保存，一周内有效。
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溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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