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| 产品编号 | 产品名称 | 产品包装 | 产品价格 |
| C2911S | AAV-Max Lysis Buffer (10X) | 100ml | 452.00元 |
| C2911M | AAV-Max Lysis Buffer (10X) | 500ml | 1898.00元 |
| C2911L | AAV-Max Lysis Buffer (10X) | 3L | 9768.00元 |
碧云天的AAV-Max Lysis Buffer (10X),中文名增强AAV收获量细胞裂解液(10X),也称Enhanced Adeno-associated Virus Cell Lysis Buffer (10X),即增强型腺相关病毒包装细胞裂解液,是一种化学成分确定(Chemical-defined, CD)的、不含任何动物源性物质(Animal origin-free, AOF)的,专用于充分裂解包装AAV用的293等细胞,以更大限度地释放AAV病毒的裂解液。本产品采用比较温和的去垢剂配方,无需额外机械破碎即可高效裂解细胞并充分释放AAV,为提取细胞中的AAV提供了一种温和、便捷且高效的方法。
腺相关病毒(Adeno-associated Virus, AAV)是Dependovirus属Parvovirus家族的微小单链DNA病毒,包含100多种血清型。AAV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基因,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,因此在生物医学领域有非常广泛的应用。
本产品操作便捷、重复性好。可以直接将本产品按照1:9的体积比加入细胞样品中,然后37ºC振荡孵育2小时,即可高效裂解细胞并释放AAV,相较于操作繁琐的反复冻融法,本产品的操作更便捷,重复性更好。
本产品提取AAV效率高。在相同条件下,使用本产品获得的AAV得率比反复冻融法要提升约115%,显著提高AAV的产率。
使用本产品、Western及IP细胞裂解液(P0013)和反复冻融法(Control)提取AAV的效果对比请参考图1。
图1.碧云天AAV-Max Lysis Buffer (10X) (C2911)、Western及IP细胞裂解液(P0013)和反复冻融法(Control)提取AAV的效果对比图。Western及IP细胞裂解液(P0013)为碧云天生产的可在非变性条件下裂解细胞的裂解液。AAV包装72小时后,收集细胞和细胞培养上清,混合均匀。三组待处理样品的起始细胞量一致,分别使用反复冻融法、P0013和本产品裂解并提取AAV,裂解的终体积均保持一致。裂解完成后,所有样品均在4ºC,13,000×g离心10分钟,收集上清,即为提取的AAV。使用碧云天的Universal AAV Titration Kit by qPCR (C2921)测定AAV滴度。结果显示,AAV-Max Lysis Buffer (10X)提取的AAV得率比对照组反复冻融法提升约115%,比Western及IP细胞裂解液(P0013)处理组提升约81%。实际检测数据会因实验条件、检测仪器等的不同而存在差异,图中数据仅供参考。
本产品不含TritonX-100,能更好维持AAV的结构完整性和感染活性[1]。
使用本产品提取的AAV样品,推荐使用碧云天的Universal AAV Titration Kit by qPCR (C2921)测定滴度。
本产品小包装、中包装和大包装分别可以用于900ml、4.5L和27L病毒包装细胞样品的裂解和提取。按照每10cm培养皿细胞样品量为10ml计算,本产品小包装、中包装和大包装分别共可以用于90、450和2700个10cm培养皿细胞样品的裂解和提取。
包装清单:| 产品编号 | 产品名称 | 包装 |
| C2911S-1 | AAV-Max Lysis Buffer (10X) | 100ml |
| C2911S-2 | Lysis Enhancer (5000X) | 200μl |
| — | 说明书 | 1份 |
| 产品编号 | 产品名称 | 包装 |
| C2911M-1 | AAV-Max Lysis Buffer (10X) | 500ml |
| C2911M-2 | Lysis Enhancer (5000X) | 1ml |
| — | 说明书 | 1份 |
| 产品编号 | 产品名称 | 包装 |
| C2911L-1 | AAV-Max Lysis Buffer (10X) | 500ml×6 |
| C2911L-2 | Lysis Enhancer (5000X) | 6ml |
| — | 说明书 | 1份 |
-20ºC保存,一年有效。AAV-Max Lysis Buffer (10X) 4ºC保存,至少6个月有效。Lysis Enhancer (5000X) 4ºC保存,至少2个月有效。
注意事项:为取得最佳的使用效果,Lysis Enhancer (5000X)须尽量避免反复冻融。可以适当分装后使用。
本产品使用前请仔细阅读附录《腺相关病毒使用安全规范》。本产品使用过程中涉及AAV,其生物安全等级为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(Standard microbiological practices)。
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。
| BSL | Agents | Practices | Primary Barriers and Safety Equipment |
Facilities (Secondary Barriers) |
| 1 | Not known to consistently cause diseases in healthy adults | Standard microbiological practices | ■ No primary barriers required. ■ PPE: laboratory coats and gloves; eye, face protection, as needed |
Laboratory bench and sink required |
| 2 | ■ Agents associated with human disease ■ Routes of transmission include percutaneous injury, ingestion, mucous membrane exposure |
BSL-1 practice plus: ■ Limited access ■ Biohazard warning signs ■ “Sharps” precautions ■ Biosafety manual defining any needed waste decontamination or medical surveillance policies |
Primary 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 needed |
BSL-1 plus: ■ Autoclave available |
| 3 | Indigenous or exotic agents that may cause serious or potentially lethal disease through the inhalation route of exposure | BSL-2 practice plus: ■ Controlled access ■ Decontamination of all waste ■ Decontamination of laboratory clothing before laundering |
Primary 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 needed |
BSL-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 exit |
| 4 | ■ 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 transmission |
BSL-3 practices plus: ■ Clothing change before entering ■ Shower on exit ■ All material decontaminated on exit from facility |
Primary barriers: ■ All procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive pressure suit |
BSL-3 plus: ■ Separate building or isolated zone ■ Dedicated supply and exhaust, vacuum, and decontamination systems ■ Other requirements outlined in the text |
| 产品编号 | 产品名称 | 包装 |
| C2901 | 通用型病毒浓缩试剂盒 | 可用于150ml/600ml/3L |
| C2906 | AAV Purification Kit Mini for All Serotypes | 5次/20次 |
| C2908 | AAV Purification Kit Midi for All Serotypes | 5次/20次 |
| C2911 | AAV-Max Lysis Buffer (10X) | 100ml/500ml/3L |
| C2912 | AAV-Max Lysis Buffer Plus (10X) | 100ml/500ml/3L |
| C2921 | Universal AAV Titration Kit by qPCR | 50次/200次 |
| C2923 | Adenovirus Titration Kit by qPCR | 50次/200次 |
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