ATF-2 Fusion Protein

• 产品编号：CST-9224S      品牌：CST       原厂货号：9224S
• 产品分类：蛋白质检测与分析 > 蛋白质，多肽和酶 > 其它纯化的蛋白及酶
• 应用分类：

描述：

ATF-2融合蛋白能够作为SAPK/JNK、 p38 MAPK的有效底物。它是一种重组融合蛋白，包含ATF-2 蛋白中的19-96残基。 其中有ATF-2 蛋白胺端的活化结构域，通过Thr69和Thr71位点的磷酸化来调节其活性。在体外激酶检测中，使ATF-2融合蛋白在反应中的浓度达到2 µg/20 µl，就能被上游激酶磷酸化，使用的试剂是1X Kinase Buffer (#9802) 和 200 µM ATP (#9804)。30ºC反应30分钟后，用免疫印迹法就能检测到磷酸化，使用的抗体是 Phospho-ATF-2 (Thr71) Antibody (#9221)。转录因子ATF-2（也被称为 CRE-BP1）能够结合到AP-1和 CRE DNA反应元件，是ATF/CREB亮氨酸拉链蛋白质家族的成员(1)。ATF-2蛋白能够调控多种病毒癌基因以及抑癌基因的表达，是SAPK/JNK 、p38 MAPK信号传导通路的靶标 (2-4)。多种细胞压力包括基因毒剂、炎性细胞因子和紫外辐射，都能激活ATF-2蛋白的转录活性。细胞应激刺激通过磷酸化Thr69和Thr71位点使ATF-2蛋白被激活 (2-4)。在体外以及转染了ATF-2的细胞中，SAPK、p38 MAPK蛋白都被证明能够磷酸化ATF-2蛋白的这些位点。这些位点的突变能导致应激性的ATF-2诱导转录活性降低(2-4)，并且降低E1A 、Rb蛋白通过ATF-2介导的基因表达。(2)。

1. Abdel-Hafiz, H.A. et al. (1992) Mol Endocrinol 6, 2079-89.
2. Gupta, S. et al. (1995) Science 267, 389-93.
3. van Dam, H. et al. (1995) EMBO J 14, 1798-811.
4. Livingstone, C. et al. (1995) EMBO J 14, 1785-97.

原厂资料：

Description

Activating Transcription Factor 2 (ATF-2) Fusion Protein serves as a useful substrate for SAPK/JNK and p38 MAP kinases. It is expressed as a recombinant protein fusion containing ATF-2 residues 19-96. It contains the amino-terminal activation domain of ATF-2, which is regulated by phosphorylation of Thr69 and Thr71.

Directions for Use

ATF-2 Fusion Protein, at a concentration of 2 µg/20 µl reaction, can be phosphorylated by an upstream kinase in an in vitro kinase assay with 1X Kinase Buffer (#9802) and 200 µM ATP (#9804). After a 30-minute assay at 30ºC, phosphorylation can be detected by Western blot with Phospho-ATF-2 (Thr71) Antibody (#9221).

Background

The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins (1). ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways (2-4). Various forms of cellular stress, including genotoxic agents, inflammatory cytokines, and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71 (2-4). Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2. Mutations of these sites result in the loss of stress-induced transcription by ATF-2 (2-4). In addition, mutations at these sites reduce the ability of E1A and Rb to stimulate gene expression via ATF-2 (2).

1. Abdel-Hafiz, H.A. et al. (1992) Mol Endocrinol 6, 2079-89.
2. Gupta, S. et al. (1995) Science 267, 389-93.
3. van Dam, H. et al. (1995) EMBO J 14, 1798-811.
4. Livingstone, C. et al. (1995) EMBO J 14, 1785-97.

注意事项：

Storage: Supplied in 20 mM Tris-HCl (pH 7.5 at 25°C), 50
mM NaCl, 2 mM Na2EDTA, 1 mM dithiothreitol (DTT) and
50% glycerol. Store at –20°C.