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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
常温,避光
- 克隆性:
单克隆
- 抗体名:
EPHA4 / HEK8 / TYRO1抗体
免疫原 :
Recombinant human EPHA4 protein ( Catalog#11314-H08H )
Antibody Type : Rabbit Polyclonal Antibody ( Antibody Purification Platform )
抗体宿主 :
Rabbit IgG
缓冲液 : 0.2 μm filtered solution in PBS, 5% trehalose may be added in some batches. Please read the hardcopy of COA or contact our customer service to confirm the formulation.
制备方法 :
Produced in rabbits immunized with purified, human cell-derived, recombinant human EPHA4 ( rh EPHA4 ; Catalog#11314-H08H ; NP_031962.2 ; Met 1 - Thr 547 ). EPHA4 specific IgG was purified by human EPHA4 affinity chromatography
EPHA4 / HEK8 / TYRO1抗体背景综述
Ephrin type-A receptor 4, also known as Tyrosine-protein kinase receptor SEK, Tyrosine-protein kinase TYRO1, EPH-like kinase 8, and EPHA4, is a single-pass type I membrane protein which belongs to the protein kinase superfamily and ephrin receptor subfamily. EPHA4 contains two fibronectin type-III domains, one protein kinase domain and one SAM (sterile alpha motif) domain. EPHA4 is a receptor for members of the ephrin-A family. EPHA4 binds to ephrin-A1, ephrin-A4 and ephrin-A5. It binds more poorly to ephrin-A2 and ephrin-A3. EPHA4 may play a role in a signal transduction process involved in hindbrain pattern formation.
The ephrins (Eph) and Eph receptors comprise the largest subfamily of receptor protein-tyrosine kinases and have been implicated in mediating developmental events, especially in the nervous system and in erythropoiesis. Based on their structures and sequence relationships, ephrins are divided into the ephrin-A (EFNA) class, which are anchored to the membrane by a glycosyl- phosphatidylinositol linkage, and the ephrin-B (EFNB) class, which are transmembrane proteins. All ephrins share a conserved extracellular sequence, which most likely corresponds to the receptor-binding domain. In molecular biology, ephrins and Eph receptors are components of cell signaling pathways involved in animal development, and implicated in some cancers. Eph receptors are classified as receptor tyrosine kinases (RTKs), and form the largest sub-family of RTKs.
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文献和实验载体克隆到腺病毒载体 -- 0-15天 4天 获得重组腺病毒 14-21天 17-22天 4-7天 噬斑纯化 5-10天 0-5天 -- 总计 26-45天 24-56天 10-17天 以体外连接方法为基础 (by Mizuguchi和Kay),CLONTECH的Adeno-X™表达系统提供了一种快速、高效构建重组腺病毒的方法。只需4至7天就可以获得重组腺病毒,如图1所示。由于经过7天的培养之后HEK 293细胞会渐渐失去对培养皿的附着而扩散,导致噬斑融合,用传统方法制备重组腺病毒往往需要铺平
(by Mizuguchi和Kay),CLONTECH的Adeno-X™表达系统提供了一种快速、高效构建重组腺病毒的方法。只需4至7天就可以获得重组腺病毒。由于经过7天的培养之后HEK 293细胞会渐渐失去对培养皿的附着而扩散,导致噬斑融合,用传统方法制备重组腺病毒往往需要铺平板以防止细胞扩散。而用CLONTECH的Adeno-X™表达系统只需4至7天即可得到纯的重组腺病毒,这意味着无需铺琼脂糖平板。Adeno-X表达系统含有一种小而便捷的穿梭载体―pShuttle,用于克隆目的基因。这种载体的表达框两侧含有极稀少
过程: 1. 铺细胞: 选择状态良好的293T细胞传代, 2-3x105个细胞/35mm dish。 2. 20-24h后,待细胞长至铺满瓶底约50-70%的时候,进行转染。下面就35mm dish为例,采用以下转染体系: ddw: 105ul plasmid: 2 ug (如0.5ug/ul, 即用4ul) 2M CaCl2: 16.5ul 2XHBS: 125ul 按上述顺序,往eppendorf管中依次加入上述四种试剂。 先将前三者混匀, 最后加2XHBS。 一种
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