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- 文献和实验
- 技术资料
- 库存：
大量
- 运输方式：
冻存运输
- 细胞形态：
成纤维样
- 物种来源：
小鼠
- 是否是肿瘤细胞：
0
- 细胞类型：
其他细胞类型
- ATCC Number：
CRL-2820™
- 生长状态：
贴壁生长
- 器官来源：
胚胎
- 年限：
14.5 days gestation embryo
| Designations: | 88TAg | ||
| Depositors: | RW Sobol | ||
| Biosafety Level: | 2 [cells containing SV40 viral DNA sequences ] | ||
| Shipped: | frozen | ||
| Medium & Serum: | See Propagation | ||
| Growth Properties: | adherent | ||
| Organism: | Mus musculus | ||
| Morphology: | fibroblast |
||
| Source: | Organ: embryo Cell Type: fibroblast immortalized with SV40 large T antigenSV40 large T antigen transfected |
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| Permits/Forms: | In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. | ||
| Isolation: | Isolation date: 2000 | ||
| Applications: | DNA repair studies | ||
| Age: | 14.5 days gestation embryo | ||
| Comments: | 88TAg (Polb null [-/-] is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb) null embryos at day 14.5 of gestation. The cell line was established by transfection with an expression vector for SV40 large T antigen [PubMed: 8538772]. The cells are transgenic for lambda LIZ (Lac I/cII). | ||
| Propagation: | ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C |
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| Subculturing: | Protocol:
Interval: Maintain cultures at a cell concentration between 2 X 10(3) and 1 X 10(5) cells/cm2. Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended Medium Renewal: Two to three times weekly |
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| Preservation: | Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
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| Related Products: | Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002 recommended serum:ATCC 30-2020 |
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| References: | 38299: Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772 88892: Sobol RW, et al. Base excision repair intermediates induce p53-independent cytotoxic and genotoxic responses. J. Biol. Chem. 278: 39951-39959, 2003. PubMed: 12882965 89413: Sobol RW, et al. Mutations associated with base excision repair deficiency and methylation-induced genotoxic stress. Proc. Natl. Acad. Sci. USA 99: 6860-6865, 2002. PubMed: 11983862 89432: Sobol RW, et al. The lyase activity of the DNA repair protein beta-polymerase protects from DNA-damage-induced cytotoxicity. Nature 405: 807-810, 2000. PubMed: 10866204 |
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文献和实验质，小分子或是金属。当融合有亲和标签的蛋白通过标签-配体作用结合再层析基质上时，通过洗杂步骤，即可去除掉其他的细胞成分。 为了洗脱所需要的蛋白质，通过改变缓冲液的条件，如pH，或通过竞争亲和标签和配体连接的方法来进行。 但怎样的纯化是较为成功的呢？仅达到所需的质量量级，如毫克级（或克级）即可满足吗？ 这些并不简单。质在蛋白纯化中同样尤为重要，比如，就蛋白的生物活性和纯度而言，通常会有所要求。 以下我们来讨论并且比较2种技术： His-tag系统和Strep-tag®系统，都使用了亲和层析法来纯化
Strep-tag®技术是亲和纯化重组蛋白的常用工具，它基于自然界中最强的非共价相互作用之一，即生物素与链霉亲和素的相互作用。该纯化系统的突出特点是纯化过程温和、目的蛋白纯度高、特异性强、兼容多种表达系统，对于有挑战性蛋白的纯化十分有帮助，因此近些年来脱颖而出。 本文将从以下方面介绍Strep-tag®技术： 标签介绍 纯化原理 标签-配体 纯化步骤 系统特点 标签介绍 目前有2种广泛使用的Strep标签： 它们分别是： Strep-tag®II 8 aa小标签(Trp-Ser
1. CUT&Tag 适用于什么物种？如果需要做植物 CUT&Tag ，需要怎么操作？ CUT&Tag 适用于哺乳动物细胞的蛋白-DNA 互作研究，酵母、植物等细胞需要经过(破除细胞壁或者提取细胞核)来进行实验。CUT&Tag 在哺乳动物细胞系上的应用比较成熟，动物组织经过处理得到悬浮单个细胞同样可以进行实验。植物进行 CUT&Tag 操作可以参照 Low-input chromatin profiling in Arabidopsis
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