| 货号 | 7880C |
| 描述 | The PathScan® Total β-Actin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of β-actin. A β-actin rabbit antibody has been coated onto the microwells. After incubation with cell lysates, β-actin is captured by the coated antibody. Following extensive washing, a pan-actin mouse detection antibody is added to detect the captured β-actin. 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 β-actin. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse/Rat/Hamster/Monkey |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total β-Actin Sandwich ELISA Kit detects endogenous levels of β-actin. As shown in Figure 1, β-actin is readily detected in HeLa cells using the PathScan® Total β-Actin Sandwich ELISA Kit. Total levels of β-actin remain unchanged after IFN-α treatment as shown by western analysis. The PathScan® Total β-Actin Sandwich ELISA Kit does not cross-react with α-smooth muscle actin, α-sarcomeric muscle actin or γ-actin. 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 | Actin, a ubiquitous eukaryotic protein, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle β- and γ-actin, also known as cytoplasmic actin, are predominantly expressed in nonmuscle cells, controlling cell structure and motility (1). α-cardiac and α-skeletal actin are expressed in striated cardiac and skeletal muscles, respectively; two smooth muscle actins, α- and γ-actin, are found primarily in vascular smooth muscle and enteric smooth muscle, respectively. These actin isoforms regulate the contractile potential of muscle cells (1). Actin exists mainly as a fibrous polymer, F-actin. In response to cytoskeletal reorganizing signals during processes such as cytokinesis, endocytosis, or stress, cofilin promotes fragmentation and depolymerization of F-actin, resulting in an increase in the monomeric globular form, G-actin (2). The Arp2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments (2). It has been reported that actin is hyperphosphorylated in primary breast tumors (3). Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle (4-6). Actin cleavage by caspase-3 may accelerate ubiquitin/proteosome dependent muscle proteolysis (6). |
| 存放说明 | 4C |
| 参考文献 | Herman, I.M. (1993) Curr. Opin. Cell Biol. 5, 48-55. Condeelis, J. (2001) Trends Cell Biol. 11, 288-293. Lim, Y.P. et al. (2004) Clin. Cancer Res. 10, 3980-3987. Kayalar, C. et al. (1996) Proc. Natl. Acad. Sci. USA. 93, 2234-2238. Communal, C. et al. (2002) Proc. Natl. Acad. Sci. USA. 99, 6252-6256. Du, J. et al. (2004) J. Clin. Invest. 113, 115-123. |
Figure 1. Treatment of HeLa cells with IFN-α stimulates phosphorylation of Stat3 at Tyr705 as detected by PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit #7300, but does not affect the level of β-actin as detected by PathScan® Total β-Actin Sandwich ELISA Kit #7880. HeLa cells were treated with 100 ng/ml IFN-α for ten minutes at 37ºC before lysis. Absorbance at 450 nm is shown in the top figure, while the corresponding western blots using Phospho-Stat3 (Tyr705) (3E2) Mouse mAb #9138 (right panel) or Total β-Actin (13E5) Rabbit mAb #4970 (left panel) are shown in the bottom figure. 图1：使用IFN-α刺激Stat3蛋白在Tyr705位点磷酸化的HeLa细胞系，随后被PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit #7205检测，但是通过PathScan® Total β-Actin Sandwich ELISA Kit #7880分析不影响β-actin蛋白水平。 HeLa细胞在裂解之前在37ºC温度下使用100 ng/ml IFN-α处理10min。在450 nm吸光度值显示在最上图中，然而通过使用Phospho-Stat3 (Tyr705) (3E2) Mouse mAb #9138 (右图)和Total β-Actin (13E5) Rabbit mAb #4970 (左图)，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 HeLa cells and the absorbance at 450 nm is shown. 图2： HeLa细胞裂解的蛋白浓度与在450 nm吸光度的关系被显示。 |
