PathScan? Phospho-ATF-2 (Thr71) Sandwich ELISA kit

• 产品编号：CST-7185C      品牌：CST       原厂货号：7185C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

CST的PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA Kit能够检测内源性Phospho-ATF-2蛋白的水平。使用Sandwich ELISA Kit #7185试剂盒能够检测出经茴香霉素处理过的 NIH/3T3细胞内的phospho-ATF-2蛋白的显著增加。而用PathScan® Total ATF-2 Sandwich ELISA Kit #7195检测总ATF-2蛋白（磷酸化和非磷酸化）水平保持不变（图1）。该试剂盒也能用于检测HeLa、293细胞中磷酸化的ATF-2蛋白。经内部测试确认，本试剂盒可以检测到所列物种中的蛋白。但对其他物种中的同源蛋白可能也可以检测。CST的PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA Kit是通过固相夹心酶联免疫吸附法（ELISA）来检测内源性的phospho-ATF-2 (Thr71) 蛋白的水平。先将Phospho-ATF-2 (Thr 71) Antibody 包被在微孔板上。与细胞裂解物孵育后，phospho-ATF-2蛋白便可被包被的抗体捕获。清洗彻底后，加入ATF-2 Mouse mAb检测phospho-ATF-2蛋白的水平。再加入Anti-mouse IgG, HRP偶联抗体#7076* 作为二抗来识别之前绑定的检测抗体。最后加入HRP底物—— TMB，进行显色。产生的吸光值与phospho-ATF-2蛋白的数量成正比。*试剂盒中抗体是专为本试剂盒特制的。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.
5. Emre, Y. et al. (2007) Biochem J 402, 271-8.
6. Ipaktchi, K. et al. (2006) J Immunol 177, 8065-71.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA Kit detects endogenous levels of Phospho-ATF-2 protein. Using this Sandwich ELISA Kit #7185, a significant induction of phospho-ATF-2 in NIH/3T3 cells treated with anisomycin can be detected. However, the level of total ATF-2 (phospho- and nonphospho-), detected by PathScan® Total ATF-2 Sandwich ELISA Kit #7195, remains unchanged (Figure 1). This kit can also be used to detect phosphorylated ATF-2 protein in human HeLa and 293 cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Phospho-ATF-2 (Thr71) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-ATF-2 (Thr71) protein. A Phospho-ATF-2 (Thr 71) Antibody has been coated onto the microwells. After incubation with cell lysates, phospho-ATF-2 protein is captured by the coated antibody. Following extensive washing, a total ATF-2 Mouse mAb is added to detect the captured phospho-ATF-2 protein. HRP-linked Anti-mouse Antibody #7076* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-ATF-2 (Thr71). 
 * Antibodies in this kit are custom formulations specific to the kit.

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.
5. Emre, Y. et al. (2007) Biochem J 402, 271-8.
6. Ipaktchi, K. et al. (2006) J Immunol 177, 8065-71.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).