| 货号 | 7385C |
| 描述 | The PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of CREB when phosphorylated at Ser133. A CREB rabbit antibody has been coated onto the microwells. After incubation with cell lysates, CREB protein (phosphorylated and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-CREB (Ser133) mouse monoclonal detection antibody is added to detect the captured phospho-CREB (Ser133) protein. HRP-linked anti-mouse IgG 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 CREB phosphorylated at Ser133. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse/Rat |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit #7385 detects endogenous levels of CREB protein when phosphorylated at Ser133 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. This kit also detects the phosphorylated form of the CREB-related protein ATF-1. |
| 供应商 | CST |
| 背景 | CREB is a bZIP transcription factor that activates target genes through cAMP response elements. CREB is able to mediate signals from numerous physiological stimuli, resulting in regulation of a broad array of cellular responses. While CREB is expressed in numerous tissues, it plays a large regulatory role in the nervous system. CREB is believed to play a key role in promoting neuronal survival, precursor proliferation, neurite outgrowth, and neuronal differentiation in certain neuronal populations (1-3). Additionally, CREB signaling is involved in learning and memory in several organisms (4-6). CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners. CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk, Ca2+, and stress signaling. Some of the kinases involved in phosphorylating CREB at Ser133 are p90RSK, MSK, CaMKIV, and MAPKAPK-2 (7-9). |
| 存放说明 | 4C and -20C |
| 参考文献 | Lonze, B.E. et al. (2002) Neuron 34, 371-85. Lee, M.M. et al. (1999) J Neurosci Res 55, 702-12. Redmond, L. et al. (2002) Neuron 34, 999-1010. Dash, P.K. et al. (1990) Nature 345, 718-21. Yin, J.C. et al. (1994) Cell 79, 49-58. Guzowski, J.F. and McGaugh, J.L. (1997) Proc Natl Acad Sci USA 94, 2693-8. Xing, J. et al. (1998) Mol Cell Biol 18, 1946-55. Ribar, T.J. et al. (2000) J Neurosci 20, RC107. Tan, Y. et al. (1996) EMBO J 15, 4629-42. |
Figure 1. Treatment of SK-N-MC cells with forskolin stimulates phosphorylation of CREB at Ser133, detected by the PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit #7385, but does not affect the levels of total CREB detected by PathScan® Total CREB Sandwich ELISA Kit #7390. SK-N-MC cells (80-90% confluent) were treated with 120 μM forskolin with 0.5 mM IBMX for 30 minutes and lysed. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using CREB (48H2) Rabbit mAb #9197 (left panel) or Phospho-CREB (Ser133) (1B6) Mouse mAb #9196 (right panel) are shown in the bottom figure. 图1：Forskolin 处理后的SK-N-MC 细胞产生了CREB 的Ser133磷酸化，为PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit #7385所检测到，但是却并不产生 PathScan® Total CREB Sandwich ELISA Kit #7390可检测到的总CREB水平的变化。SK-N-MC 细胞 (80-90% 铺满)用120 μM forskolin with 0.5 mM IBMX 处理30分钟，然后裂解。OD 450 读数在上图显示，对应的使用CREB (48H2) Rabbit mAb 兔单抗 #9197（左）或Phospho-CREB (Ser133) (1B6) Mouse mAb 鼠单抗#9196（右）的结果在下图显示。 | |
Figure 2. The relationship between the protein concentration of lysates from untreated and forskolin-treated SK-N-MC cells and the absorbance at 450 nm using the PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit is shown. 图2：显示未处理和经forskolin处理的SK-N-MC 细胞提取液中的蛋白浓度与450 nm 吸光值的关系。使用的试剂盒是 PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit。 |
