Recombinant Human NRas (Flag-Tag)
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产品编号 P2401-100μg
数量
¥ 2698.00
产品包装:
10μg 100μg 1mg
产品简介
使用说明
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Species Gene ID Accession Source Length MW Tag
Human 4893 P01111 E.coli 196aa ~22kDa N-Flag Tag

蛋白信息(About this protein)
名称(Name) Recombinant Human NRas; 重组人NRas蛋白
别名(Synonyms) Recombinant Human N-Ras, 重组人N-Ras蛋白, NRas Proto-Oncogene; Transforming Protein N-Ras; Neuroblastoma Ras Viral (V-Ras) Oncogene Homolog, Neuroblastoma Ras Viral oncogene Homolog; proto-oncogene GTPase; ALPS4; AV095280; N-Ras; HRAS1; NS6
产品简介(Background) Beyotime’s recombinant human NRas (rhNRas) was expressed in E.coli and purified, which contain the mature form NRas (2-186aa) with Flag tag (DYKDDDDK) at the N-terminus.
Ras (NRas, HRas and KRas) is the most frequently mutated gene family in cancers,they differ significantly only in the C-terminal 40 amino acids. These Ras genes have GTP/GDP binding and GTPase activity, and their normal function may be as G-like regulatory proteins involved in the normal control of cell growth. Ras directly interacts with and activates several downstream effector pathways including the mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways. Mutations in Ras gene disrupt the guanine exchange cycle, typically by becoming GAP-independent and ‘locking’ Ras in the active, GTP-bound state, thereby activating downstream signaling pathways resulting in tumor cell growth.
NRas oncogene encoding a membrane protein that shuttles between the Golgi apparatus and the plasma membrane. This shuttling is regulated through palmitoylation and depalmitoylation by the ZDHHC9-GOLGA7 complex. Mutations which change amino acid residues 12, 13 or 61 activate the potential of NRas to transform cultured cells and are implicated in a variety of human tumors, such as somatic rectal cancer, follicular thyroid cancer, autoimmune lymphoproliferative syndrome, Noonan syndrome, and juvenile myelomonocytic leukemia [1].
产品用途(Applications) Recombinant Human NRas is useful in studying NRas interacting proteins, effectors, GAPs (GTPase-activating proteins) and GEFs (Guanine nucleotide-exchange factors). It can also be used as positive control in Western blots.
外观(Physical appearance) Liquid
活性(Biological activity) The specific activity of NRas was 4.3 nmol/min/mg in GTPase activity assay.
浓度(Concentration) 1mg/ml
纯度(Purity) ≥ 95% by SDS-PAGE
储存液(Storage buffer) 20mM HEPES (pH7.4), 200mM NaCl, 1mM EDTA, 2mM DTT, 5% glycerol

小GTP酶(Small GTPase),也称Small G-proteins、Ras superfamily,是调节真核细胞信号转导、细胞增殖、细胞骨架重组和细胞内膜转运等过程的分子开关,小GTP酶通过结合和水解GTP,在“激活”和“静息”状态之间循环:在外界信号的刺激下,鸟苷酸交换因子(Guanine nucleotide-exchange factors, GEFs)辅助小GTP酶将结合的GDP置换为GTP,小GTP酶结合GTP进入激活状态(Active state);激活状态的小GTP酶与下游效应蛋白(Effector protein)相互作用,从而刺激细胞发生相应的响应;GTP酶激活蛋白(GTPase-activating proteins, GAPs)催化小GTP酶结合的GTP水解为GDP,并释放自由磷酸盐(Free phosphate, Pi),此时小GTP酶结合GDP进入静息状态(Inactive state),鸟嘌呤核苷酸解离抑制蛋白(Guanine nucleotide dissociation inhibitors, GDIs)抑制小GTP酶释放GDP,直到GEFs受到刺激信号再次开启新一轮的循环(图1) [2-3]。

图1.小GTP酶在“激活”和“静息”状态之间循环的原理图。

小GTP酶在人类中已发现超过150个家族成员,在果蝇(Drosophila)、秀丽隐杆线虫(C. elegans)、酿酒酵母(S. cerevisiae)、粟酒裂殖酵母(S. pombe)、黏菌(Dictyostelium)和植物中也都发现了保守的同源物。小GTP酶根据结构和功能被分为5个家族分支:Ras家族、Rho家族、Ran家族、Rab家族和Arf家族。Ras家族本身又被分为6个亚家族:Ras亚家族、Ral亚家族、Rit亚家族、Rap亚家族、Rheb亚家族和Rad亚家族[4]。

Ras家族(Ras homologous GTPases)是调节肌动蛋白重组的关键因子,因此在肌动蛋白细胞骨架完整性、细胞增殖、细胞分化、细胞粘附、细胞凋亡和细胞迁移等细胞过程中发挥着重要作用。目前有16个成员被发现,被分为6个亚家族(见下表),其中NRas、HRas和KRas研究的最为广泛[5]。

Subfamily Ras Ral Rit Rap Rheb Rad
Subfamily members HRas
NRas
KRas4A
KRas4B
RalA
RalB
Rit1
Rit2
Rap1A
Rap1B
Rap2A
Rap2B
Rap2C
Rheb
RhebL1
RRad

本产品经SDS-PAGE电泳检测蛋白纯度和分子量参考图2。

图2.碧云天Recombinant Human NRas (P2401)的SDS-PAGE电泳检测图。本蛋白经BeyoGel™ Plus PAGE预制胶(Tris-Gly, 4-15%, 10孔) (P0465)电泳,Marker为BeyoColor™彩色预染蛋白分子量标准(6.5-270kD) (P0071/P0072),并经BeyoBlue™考马斯亮蓝超快染色液(P0017F)染色。实际检测结果可能会因样品和检测条件等的不同而存在差异,图中数据仅供参考。

包装清单:
产品编号 产品名称 包装
P2401-10μg Recombinant Human NRas (Flag-Tag) 10μg
P2401-100μg Recombinant Human NRas (Flag-Tag) 100μg
P2401-1mg Recombinant Human NRas (Flag-Tag) 1mg
说明书 1份
保存条件:

-20ºC保存,一年有效。-80ºC保存,可以保存更长时间。

注意事项:

由于蛋白每次冻融均会引起部分失活,所以首次配制成相应浓度的储存液后,须分装后-20ºC或更低温度冻存,尽量避免反复冻融。

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.收到产品后请立即按照说明书推荐的条件保存。在打开管盖前,请适当离心,使附着在管盖或管壁上的蛋白聚集于管底。
2.具体的最佳工作浓度请自行参考相关文献,或者根据实验目的,通过实验进行摸索和优化。
参考文献:
1.Moore AR, Rosenberg SC, McCormick F, Malek S. Nat Rev Drug Discov. 2020. 19(8):533-52.
2.Vernoud V, Horton AC, Yang Z, Nielsen E. Plant Physiol. 2003. 131(3):1191-208.
3.Ito H, Morishita R, Nagata KI. Int J Mol Sci. 2018. 19(7):2121.
4.Goitre L, Trapani E, Trabalzini L, Retta SF. Methods Mol Biol. 2014. 1120:1-18.
5.Bos JL. Cancer Research. 1989. 49(17): 4682–9.
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