| 货号 | 7249C |
| 描述 | The PathScan® Phospho-YB1 (Ser102) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of YB1 when phosphorylated at Ser102. A YB1 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, YB1 protein is captured by the coated antibody. Following extensive washing, a phospho-YB1 (Ser102) mouse detection antibody is added to detect the captured phospho-YB1 (Ser102) protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of YB1 phosphorylated at Ser102. Antibodies in kit are custom formulations specific to kit.The PathScan ® Phospho-YB1 (Ser102) Sandwich ELISA Kit是一种固相三明治酶联免疫吸附实验（ELISA），用来检测在Ser102 发生磷酸化的内源性YB1。YB1兔抗体已经包被在孔上，与细胞裂解物共孵育后，YB1蛋白被报备的抗体捕获。经过彻底清洗，加入磷酸化YB1 （ Ser102 ）抗体检测被捕获的phospho-YB1 (Ser102)蛋白。抗鼠IgG，HRP直标抗体用来识别结合的检测抗体。HRP底物TMB加入后产生颜色。颜色的吸光度值与Ser102磷酸化的YB1蛋白的量呈正相关。本试剂盒中的抗体是本试剂盒的特定抗体。 |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-YB1 (Ser102) Sandwich ELISA Kit #7249 detects endogenous levels of YB1 protein when phosphorylated at Ser102 as shown in Figure 1. Kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 供应商 | CST |
| 背景 | The Y-box binding protein 1 (YB1) belongs to a family of evolutionarily conserved, multifunctional Y-box proteins that bind single-stranded DNA and RNA and function as regulators of transcription, RNA metabolism, and protein synthesis (1). YB1 binds to Y-box sequences (TAACC) found in multiple gene promoters and can positively or negatively regulate transcription. YB1 activates genes associated with proliferation and cancer, such as cyclin A, cyclin B1, matrix metalloproteinase-2 (MMP-2), and the multi-drug resistance 1 (MDR1) gene (2-4). YB1 represses genes associated with cell death, including the Fas cell death-associated receptor and the p53 tumor suppressor gene (5-7). It also interacts with the RNA-splicing factor SRp30c and stabilizes interleukin-2 (IL-2) mRNA upon induction of T lymphocytes by IL-2 (8,9). The majority of YB1 protein localizes to the cytoplasm, with a minor pool found in the nucleus; however, nuclear localization appears to be critical for its role in promoting proliferation. Nuclear translocation is cell cycle regulated, with YB1 protein accumulating in the nucleus during G1/S phase (2). In addition, nuclear translocation is induced in response to extracellular stimuli such as hyperthermia and UV irradiation, or treatment of cells with thrombin, interferons, or insulin-like growth factor (IGF-I) (2,10). Treatment of the MCF7 breast cancer cell line with IGF-I results in Akt-mediated phosphorylation of YB1 at Ser102, which is required for nuclear translocation of YB1 and its ability to promote anchorage-independent growth (10). Research studies have shown that YB1 is overexpressed in many malignant tissues, including breast cancer, non-small cell lung carcinoma, ovarian adenocarcinomas, human osteosarcomas, colorectal carcinomas, and malignant melanomas. Investigators have shown that nuclear YB1 expression correlates with high levels of proliferation, drug resistance, and poor tumor prognosis (2,7,10).Y-box结合蛋白1（YB1）属于一类进化上保守的，多功能Y-box蛋白，和单链DNA和RNA结合，调控转录，RNA代谢和蛋白质的合成（1）。YB1与多种基因启动子中的Y-box序列（TAACC）结合，并能正向或负向调节转录。YB1激活与增殖和癌症相关的基因，如周期蛋白A，B1，基质金属蛋白酶-2（MMP-2），及多药耐药性1（MDR1）基因（2-4）。YB1抑制与细胞死亡有关的基因，包括细胞的Fas相关死亡受体和p53抑癌基因（5-7）。它还与RNA剪切因子SRp30c互作并且能够稳定诱导T淋巴细胞的白细胞介素2的mRNA（8,9）。YB1蛋白主要定位于细胞质，少量存在于细胞核中；然而，核里面的YB1对促进细胞增殖十分关键。核转运受到细胞周期调控，在G1 / S期YB1蛋白质积累在细胞核中（2）。此外，细胞受到外部刺激，如高温和紫外线照射，或着用凝血酶，干扰素，胰岛素样生长因子（IGF-1）处理后可以诱导发生核转运（2,10）。IGF-1处理MCF7乳腺癌细胞株诱发Akt介导的YB1（102位丝氨酸）磷酸化，促进YB1的核转运及非贴壁依赖性生长（10）。YB1在许多恶性肿瘤组织中高表达，包括乳腺癌，非小细胞肺癌，卵巢癌，人骨肉瘤，大肠癌，恶性黑色素瘤。核YB1表达与过度增殖，耐药性，及肿瘤的不良预后相关（2,7,10）。 |
| 存放说明 | 4C |
| 参考文献 | Matsumoto, K. and Wolffe, A.P. (1998) Trends Cell Biol. 8, 318-23. Jurchott, K. et al. (2003) J. Biol. Chem. 278, 27988-96. Mertens, P.R. et al. (1997) J. Biol. Chem. 272, 22905-12. Uchiumi, T. et al. (1993) Cell Growth Differ. 4, 147-57. Lasham, A. et al. (2000) Gene 252, 1-13. Lasham, A. et al. (2003) J. Biol. Chem. 278, 35516-23. Homer, C. et al. (2005) Oncogene 24, 8314-25. Raffetseder, U. et al. (2003) J. Biol. Chem. 278, 18241-8. Chen, C.Y. et al. (2000) Genes Dev. 14, 1236-48. Sutherland, B.W. et al. (2005) Oncogene 24, 4281-92. |
Figure 2. The relationship between the protein concentration of lysates from Jurkat cells, treated with LY294002 (as control) or calyculin A/pervanadate, and the absorbance at 450 nm using the PathScan® Phospho-YB1 (Ser102) Sandwich ELISA Kit is shown.采用LY294002 (对照组) 和花萼海绵诱癌素A和过钒酸盐处理Jurket细胞提取物中蛋白浓度之间的关系，所用试剂盒为PathScan ® Phospho-YB1 (Ser102) Sandwich ELISA Kit ，在450nm处读取吸光度值。 | |
Figure 1. Treatment of Jurkat cells with calyculin A and pervanadate stimulates phosphorylation of YB1 at Ser102, detected by the PathScan® Phospho-YB1 (Ser102) Sandwich ELISA Kit #7249, but does not affect the levels of total YB1 detected by western. Jurkat cells were treated with 50 μM LY294002 (as control), or 100 nM calyculin A and 1 mM pervanadate for 30 minutes and lysed. The absorbance readings at 450 nm are shown in the upper figure, while the corresponding western blots using YB1 (D299) Antibody #4202 (left panel) or Phospho-YB1 (Ser102) (C34A2) Rabbit mAb #2900 (right panel) are shown in the lower figure.用花萼海绵诱癌素A和过钒酸盐处理Jurket细胞，是YB1蛋白在Ser102发生磷酸化，所用试剂盒为PathScan ® Phospho-YB1 (Ser102) Sandwich ELISA Kit #7249 ，但western检测后不影响总YB1蛋白的水平。用50 μM LY294002 (作为对照)或100 nM花萼海绵诱癌素A 和 1 mM过钒酸盐处理Juerkt细胞30分钟，然后裂解。上图数据显示在450 nm读取吸光值，相对应的western blots 结果在下图数据中，所用抗体为YB1 (D299) Antibody #4202 (左泳道) 或 Phospho-YB1 (Ser102) (C34A2) Rabbit mAb #2900 (右泳道) |
