| 货号 | 7863C |
| 描述 | The PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated DDR1 protein. A DDR1 rabbit antibody has been coated on the microwells. After incubation with cell lysates, DDR1 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse mAb is added to detect captured tyrosine-phosphorylated DDR1 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 this developed color is proportional to the quantity of DDR1 protein phosphorylated on tyrosine residues. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit detects endogenous levels of tyrosine-phosphorylated DDR1 protein in human cells, as shown in Figure 1. The 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 discoidin domain receptors (DDRs) are receptor tyrosine kinases with a discoidin homology repeat in their extracellular domains, activated by binding to extracellular matrix collagens. So far, two mammalian DDRs have been identified: DDR1 and DDR2 (1). They are widely expressed in human tissues and may have roles in smooth muscle cell-mediated collagen remodeling (2). Aberrant expression and signaling of DDRs have been implicated in human disease related to increased matrix degradation and remodeling, such as cardiovascular disease, liver fibrosis, and tumor invasion (1). |
| 存放说明 | 4C |
| 参考文献 | Vogel, W. (1999) FASEB J 13 Suppl, S77-82. Ferri, N. et al. (2004) Am J Pathol 164, 1575-85. |
Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. Calu-3 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate, respectively).图2：磷酸化或非磷酸化的裂解中蛋白含量和450nm吸光值之间的关系。Calu-3细胞在85%满时收样，并用加入或未加磷酸酶抑制剂的裂解液进行处理（磷酸化或非磷酸化）。 | |
Figure 1: Constitutive phosphorylation of DDR1 in Calu-3 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit #7863 (upper, right). In contrast, a low level of phospho-DDR1 protein is detected in Calu-3 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total DDR1 protein from both nonphospho and phospho lysates are detected by PathScan® Total DDR1 Sandwich ELISA Kit #7845 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a Phospho-DDR1 (Tyr792/796/797) rabbit antibody (right) or a total DDR1 rabbit antibody (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4.图1：在磷酸酶抑制剂*存在时（磷酸化的裂解液），用PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit #7863能够检测到Calu-3细胞中磷酸化DDR1的组成型表达（上，右）。相比之下，没有磷酸酶抑制剂*存在时（非磷酸化的裂解液），仅能检测到低水平的磷酸化DDR1的表达。用PathScan® Total DDR1 Sandwich ELISA Kit #7845则能在磷酸化和非磷酸化的裂解液中检测到近似水平的总DDR1的表达（上，左）。上图显示450nm处的吸光值，下图显示用抗磷酸化DDR1 (Tyr792/796/797)的兔源单克隆抗体（右）或抗总DDR1的兔源单克隆抗体（左）进行免疫印迹检测的结果。*磷酸酶抑制剂中包含sodium pyrophosphate、β-glycerophosphate和 Na3VO4。 |
