| 货号 | 7944C |
| 描述 | The PathScan® Total α-Tubulin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of α-tubulin. An α-tubulin rabbit antibody has been coated onto the microwells. After incubation with cell lysates, α-tubulin is captured by the coated antibody. Following extensive washing, an α-tubulin mouse detection antibody is added to detect the captured α-tubulin. An 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 α-tubulin. |
| 反应种属 | Human/Mouse/Monkey |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total α-Tubulin Sandwich ELISA Kit detects endogenous levels of α-tubulin protein, as shown in Figure 1. The kit sensitivity is shown in Figure 2. Microtubule stabilizing or destabilizing agents may significantly increase or decrease the signal, respectively. 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 cytoskeleton consists of three types of cytosolic fibers: microtubules, microfilaments (actin filaments), and intermediate filaments. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. γ-tubulin is required to nucleate polymerization of tubulin subunits to form microtubule polymers. Many cell movements are mediated by microtubule action, including the beating of cilia and flagella, cytoplasmic transport of membrane vesicles, chromosome alignment during meiosis/mitosis, and nerve-cell axon migration. These movements result from competitive microtubule polymerization and depolymerization or through the actions of microtubule motor proteins (1). |
| 存放说明 | 4C |
| 参考文献 | Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47. |
Figure 1. Treatment of A-431 cells with EGF stimulates phosphorylation of the EGF receptor at Tyr1086 as detected by PathScan® Phospho-EGF Receptor (Tyr1086) Sandwich ELISA Kit #7240, but does not affect the level of α-tubulin as detected by PathScan® Total α-Tubulin Sandwich ELISA Kit #7944. A-431 cells were treated with EGF (100 ng/ml, 3 min.) at 37ºC before lysis. Absorbance at 450 nm is shown in the top figure, while the corresponding western blots using Total α-Tubulin (11H10) Rabbit mAb #2125 (left) or Phospho-EGF Receptor (Tyr1086) Antibody #2220 (right) are shown in the bottom figure. 图1：使用EGF刺激EGF受体在Tyr1086位点磷酸化的A-431细胞系，随后被PathScan® Phospho-EGF Receptor (Tyr1086) Sandwich ELISA Kit #7240检测，但是通过PathScan® Total α-Tubulin Sandwich ELISA Kit #7944分析不影响α-tubulin蛋白水平。 A-431细胞在裂解之前在37ºC温度下使用EGF (100 ng/ml, 3 min.)处理。在450 nm吸光度值显示在最上图中，然而通过使用α-Tubulin (11H10) Rabbit mAb #2125 (左图)和Phospho-EGF Receptor (Tyr1086) Antibody #2220 (右图)，相应的免疫印迹（western blot）显示在下图中。 | |
Figure 2. The relationship between the protein concentration of the lysate from A-431 cells and the absorbance at 450 nm is shown. 图2： A-431细胞裂解的蛋白浓度与在450 nm吸光度的关系被显示。 |
